Blood component deposition-preventing agent, methods using them, and blood test ware and matrixes

ABSTRACT

This invention has for its object to provide a blood component deposition-preventing agent and a blood coagulation accelerator, which are substantially indifferent to blood coagulation activity and serum chemistry parameters and a plastic blood test ware and a blood test matrix which do not confound measured values. The invention relates to a blood component deposition-preventing agent comprising a random copolymer of a monomer component (a) giving a water-soluble homopolymer and a monomer component (b) giving a water-insoluble homopolymer, a blood coagulation accelerator comprising a substantially blood-insoluble antimicrobial composition comprising a carrier and, as supported thereon, an antimicrobial metal, and a blood test ware or matrix carrying them on its inside wall or surface.

This application is a divisional of Ser. No. 08/648,132 filed Jul. 5,1996, now U.S. Pat. No. 5,888,824 which is the national filing ofPCT/JP/95/00461, filed Mar. 17, 1995.

TECHNICAL FIELD

The present invention relates to a blood component deposition-preventingagent and a blood coagulation accelerator for use in the laboratoryexamination of a blood sample, particularly in hematology, serumbiochemistry, and immunoserology, methods using them, and bloodexamination ware and matrixes.

BACKGROUND TECHNOLOGY

With recent advances in testing techniques, the chemical,immunoserological and hematological examinations of blood have witnesseda remarkable mechanization so that it is by now certain that only ifproperly prepared samples were provided, such examinations could becarried through in short periods of time. For example, even in theoutpatient setting, the doctor would be able to make a diagnosis basedon blood examination data, thus contributing much to the diagnosis andtherapy of diseases,

As to the pretreatment for hematological examinations using whole bloodas a sample, mere admixing of the blood with an anticoagulant, which isnot time-consuming, is sufficient so that the sample can be almostimmediately set in an analyzer.

However, in biochemical or immunoserological examinations using theserum fraction of blood, it is necessary to coagulate the blood onceand, then, separate the serum by centrifugation or the like and theprocedure is rather time-consuming. Therefore, in order to reduce thetime required for the whole examination procedure from the pretreatmentof a sample to the output of tent data, mere shortening of the analysistime by mechanization of the analytical procedure is insufficient and itis necessary to shorten the time required for separation of serum.

Meanwhile, glassware has heretofore been used as the blood examinationvessel for accommodating the blood to be test, allowing it to coagulatetherein, and separating the serum by centrifugation. However, glasswareis vulnerable to mechanical impact and, when it is broken the testsample that issues out or splashes may cause the examiner to be infectedby pathogenic bacteria and, as an additional problem, the necessaryblood sampling for examination adds to a burden on the patient. Forthese reasons, plastic vessels have come into popular use in recentyears. However, such plastic ware has been found disadvantageous in thatthe formed elements of the blood (hereinafter referred to as bloodcomponents) such as platelets, various blood proteins, and especiallythe fibrins which are formed in the final stage of the blood coagulationprocess, are very liable to deposit on the inside wall of the plasticware and thereby exert untoward effects on examination results.Moreover, as will be described in detail hereinafter, it is commonpractice to employ a mineral substance or an organic substance such asellagic acid, as a blood coagulation accelerator in blood examinationware but such blood coaglation accelerator tends to encourage depositionof said blood components on the vessel wall and the blood componentsonce deposited will not easily be detached from the inside wall of thevessel under the routine conditions of centrifugation such as about 1000to 1800 G.times.5 minutes. As a result owing to the high shear force ofcentrifugation acting on the interface between the inside wall and theclot the platelets and red blood cells are destroyed and their contentsleak out to affect the examination results.

To overcome these disadvantages, Japanese Kokai PublicationSho-58-105063 and Japanese Kokai Publication Sho-58-105064 proposed amethod which comprises disposing a blood coagulation accelerator and anonionic subfactant concomitantly on the inside wall of the ware, forinstance. However, with the recent rapid development of high sensitivitytechniques in the field of immunoserological examination, analogues ofnonionic surfactants are being used as sensitizers on more and moreoccasions. If the test serum is contaminated with a nonionic surfactant,oversensitizing reactions occur in immunoserologic parameters to presentthe problem of inaccuracy leading to false positive tests.

On the other hand, for efficient separation of plasma from the blood tobe analyzed, there is a protocol involving addition of a bloodanticoagulant such an ethylenediamineteraacetic acid salt or a citrateto the blood sample. In hematological examinations using ananticoagulant, too, the deposition of blood components, particuarlyplatelets, on the plastic surface may be a cause of trouble, althoughthe frequency of the trouble is not high. If platelets stick to theinside wall of blood test ware, the platelet count may show onabnormally low value or confound blood coagulation function values.Moreover, where an emergency chemical examination is required, it iscommon practice to use a heparin salt which is a kind of anticoagulantbut if the deposition of platelets occurs in such cases, various enzymesof platelet origin leak out into the plasma with time so that therelated examination parameters tend to show abnormally high values.These events are less frequent as compared with the coagulation of bloodand have so far attracted little attention but are now pointed out as aserious problem as an omnibus, accurate and rapid blood examination isdemanded.

The above problem of deposition of blood components has been pointed outwith reference to plastic blood test ware but recently the adverseinfluences of the deposition and activation of platelets on examinationresults have been pointed out for blood examination glassware as welland improvements are being sought just as for plastic ware.

Since plastic ware for blood examination is intrinsically low in thepotential to activate blood coagulation XII factor and XI factor, ittakes by far a longer time for the blood to coagulate in plastic warethan in glassware and, therefore, plastic ware has so far been of lowpractical value.

Therefore, attempts have been made to shorten the blood coagulation timeby coating the inside wall of blood test ware with a finely dividedmineral substance such as glass, kaolin, bentonite, silica, cerite, orthe like or a blood coagulation accelerator such an ellagic acid astaught in Japanese Kokai Publication Sho-58-195151 or accommodating inthe ware a substantially blood-insoluble and chemically inert nonwovencloth or plastic sheet matrix on Which said finely divided particleshave been imobilized as taught in Japanese Kokal PublicationSho-58-105064.

When a blood coagulation-accelerating substance is to be coated on theinside wall of a blood test ware or immobilized on a carrier, asuspension of finely divided particles of such substance either in purewater or in a mixture of alcohol and pure water is prepared andspray-coated on the inner surface of the ware or a carrier material isdipped in such a suspension, dried, cut to size, and accommodated withinthe blood test ware.

However, such a treating suspension is susceptible to the attack ofmicroorganisms and unless it is properly handled, may causecontamination of a blood sample with microorganisms. Furthermore, when awater-soluble macromolecular compound such as polyvinylpyrrolidone or amodified cellulose is incorporated in said suspension as a binder forsaid coagulation accelerator powder or a viscosity adjusting agent forthe suspension as is generally practiced, the water-solublemacromolecular compound serves as a good nutrient source formicroorganisms so that the above-mentioned tendency of the treatingsuspension to be a microbial contamination risk factor in furtherencouraged.

As the proliferation of microorganisms progresses, condensation productswill be accumulated in the treating suspension to cause troubles such aclogging of the spray nozzle, marked loan of coated surface evenness,and biases in the density of particles immobilized on the carrier in thedipping stage, all of which add up to measurement errors. The risk ofmicrobial contamination is not confined to the current risk associatedwith degradation of the treating suspension but is a persistent drawbackfor the shelf-life of the ware unless the method for storage of the wareis wholesome.

Therefore, unless manufactured in a sterile environment, a blood testware containing a blood coagulation accelerator may have to besterilized with actinic radiation such as gamma-rays, electron beams,etc. or a chemically reactive gas such as ethylene oxide gas. In any ofsuch procedures the radiation dose or the concentration of the reactivegas, heating temperature, exposure time and other sterilizing conditionsmust be adjusted according to the contamination status prior tosterilization and, thus, very delicate control is required.

Moreover, where the sterilizing load has been severely contaminated,rugged sterilizing conditions are required so that the load may sustainirreversible modification, deformation, and other damages. Moreover, inthe storage after sterilization, the sterility once established will bejeopardized unless the ware is properly packaged.

One of the possible effective approaches to solving the above problemsis to impart antimicrobial activity to the very coagulation accelerator.By such a technique, the above-mentioned microbial contamination wouldbe inhibited and even if sterilization be needed, mild sterilizingconditions would be sufficient, with the result that the physical andchemical changes of the load due to sterilization could be prevented orsuppressed. Furthermore, the packaging of the blood test ware could besimplified.

As regards antibacterial and antifungal agents which are generally usedfor prevention of microbial contamination, a large number of compoundsinclusive of those for food use are already known and in use. However,the large majority of these antibacterial and antifungal agents arewater-soluble and, therefore, if the blood is drawn into a blood testware in which a blood anticoagulant supplemented with such anantimicrobial agent has been accommodated, the antimicrobial agent maydissolve out into the blood to confound various chemical tests.Moreover, when the antibacterial or antifungal agent is a water-solubleheavy metal salt, it modifies the enzymes associated with bloodcoagulation and the resulting deactivation of the enzymes preventcoagulation of the blood and make it difficult to achieve the objectivesuch as separation of serum.

Where the specimen to be analyzed is plasma, it is a routine procedureto mix the blood with an anticoagulant and centrifuge the mixture toseparate plasma from the solid fraction. Generally speaking, in order toavoid contamination of plasma with substances liberated from formedelements of the blood and other matter and the consequent interferencewith tests, the plasma obtained by centrifuging blood in the abovemanner is transferred to a different container and stored. Recently,however, for the purpose of protecting the examiner against infectionvia the patient's blood, a procedure which does not require a transferto another ware is demanded. Therefore, the use of a plasma separatorcomprising a thixotropic fluid as disclosed in Japanese KokaiPublication Hei-2-168159 or the use of a separator for provision of apartition between the plasma layer and the solid component layer astaught in Japanese Kokai Publication Hei-5-26873 has been recentlyemployed.

However, the inside surface of the plastic ware is hydrophobic and theblood cells and proteins are adsorbed thereon as mentioned above.Particularly platelets are adsorbed with a high affinity and because LDD(lactic dehydrogenase), CPK (creatine kinase), K (potassium), etc. occurat higher levels in platelets than in plasma, these components arereleased from the platelets adsorbed on the plastic surface and it isinevitable that the values of these test parameters are considerablyaffected.

Therefore, even if the above-mentioned procedure of providing apartition between the plasma layer and the solid component layer isfollowed, the gradual release of enzymes and others from the blood cellsadsorbed on the inside wall of the ware containing the plasma isunavoidable and interferes with tests. These adverse effects areparticularly remarkable when the plasma is stored in the refrigeratorfor reexamination.

Having overcome the above disadvantages of the prior art, the presentinvention has for its primary object to provide a blood componentdeposition-preventing agent which is capable of inhibiting deposition ofblood components effectively without causing the problem of falsepositive reactions in the immunoserological tests.

The second object of the present invention is to provide a bloodcoagulation accelerator comprising an antibacterial composition whichhas its own blood coagulation-accelerating activity and yetsubstantially does not interfere with blood coagulation activity orconfound serum biochemical tests.

The third object of the present invention is to provide a plastic wareand a matrix for blood examination which do not influence test valuesowing to release of substances from blood cells even when used in testson plasma.

DISCLOSURE OF INVENTION

The essential feature of the first aspect of the present invention isthat a random copolymer comprising 10 to 90 mol % of a monomer component(a) the homopolymer of which is water-soluble and 90 to 10 mol % of amonomer component (b) the homopolymer of which in water-insoluble isused as a blood component deposition-preventing agent.

The essential feature of the second aspect of the present invention isthat a blood coagulation accelerator is provided by supporting anantimicrobial metal on a carrier material and incorporating theresulting substantially blood-insoluble antimicrobial composition.

The essential feature of additional aspect of the present inventionwhich is composed of first and second aspects resides in methods usingsaid blood component deposition-preventing agent and said bloodcoagulation accelerator and in the blood test ware and matrix.

The first and second aspects of the present invention are now describedin detail.

The blood component deposition-preventing agent according to the firstaspect of the present invention comprises a random copolymer. Themonomer component(a) as a constituent of said random copolymer is notlimited in kind only if its homopolymer is water-soluble,thus includingvinylpyrrolidone, vinyl alcohol, ethylene oxide, salts of acrylic acid,salts of styrenesulfonic acid, salts of vinyl phosphonic acid,allylamine salts, hydroxymethyl(meth)acrylate,glycosylethyl(meth)acrylate, saccharides such as glucose, amino acidssuch as glutamic acid, and so on. These monomers can be used singly orin combination.

The monomer component (b) as the other constituent of said randomcopolymer is not limited in kind, either, only if its homopolymer iswater-insoluble, thus including ethylene, propylene, propylene oxide,vinyl acetate, vinyl chloride, alkyl(meth)acrylates, styrene,acrylonitrile, acrolein, and so on. These monomers can be used singly orin combination.

The random copolymer comprising said monomer component (a) and monomercomponent (b) can be provided by the known addition polymerization,polycondensation, or other technique.

However, from the standpoint of availability, it is advantageous toprepare the random copolymer using vinylpyrrolidone or vinyl alcohol asmonomer component (a) and vinyl acetate as monomer component (b). Ascommercial products, Luviskol VA grade numbers VA73, VA64, VA55, VA37,and VA28 are available from BASF as typical vinylpyrrolidone-vinylacetate random copolymers and Unitika Poval grade numbers E-180,UMR-10M, UMR-30L, and UMR-150L are available from Unitika Ltd. astypical vinyl alcohol-vinyl acetate random copolymers.

In the above random copolymer, the proportion of monomer component (a)the homopolymer of which is water-soluble is within the range of 10 to90 mol % and that of monomer component (b) the homopolymer of which iswater-insoluble is within the range of 90 to 10 mol %.

If the proportion of monomer component (a) the homopolymer of which iswater-soluble exceeds 90 mol %, the resulting random copolymer will notbe much different in characteristics from the homopolymer of monomercomponent (a) so that the rate of adsorption on the inside wall of bloodtest ware and on the matrix surface is decreased and the solubility inblood is too high. As a consequence, when blood is drawn into the bloodteat ware, the random copolymer is washed out from the inner surface ofthe ware and the matrix surface so that it cannot play the role ofpreventing deposition of blood components.

On the other hand, if the proportion of monomer component (a) thehomopolymer of which is water-soluble is less than 10 mol %, thecopolymer will not be much different in characteristics from thehomopolymer of monomer component (b) and become substantially insolublein blood so that it may not play the role of preventing deposition ofblood components. When a combination of this blood componentdeposition-preventing agent and a blood coagulation accelerator or ablood anticoagulant is applied to the inside wall of the blood test wareor the matrix surface and dried, a blood-insoluble film is formed on thesurface of the blood coagulation accelerator or anticoagulant so thatblood coagulation factors XII, XI, etc. cannot bind to the surface ofthe blood coagulation accelerator, with the result that the coagulationof blood is not hastened and the reduced solubility of the bloodanticoagulant leads to an insufficient anticoagulant effect.

For the above reasons, the random copolymer for use in the presentinvention should be such that said monomer component (a) whosehomopolymer is water-soluble and said monomer component (b) whosehomopolymer is water-insoluble account for 10 to 90 mol % and 90 to 10mol %, respectively.

The blood test ware of the present invention comprises a vessel and, asdisposed on its inner surface, the blood component deposition-preventingagent according to the first aspect of the present invention.

In this construction, the amount of the blood componentdeposition-preventing agent present on the inner surface of the vesselis preferably in the range of 1×10⁻¹⁰ to 1×10⁻² g/cm². If the amount ofthe blood component deposition-preventing agent is less than 1×10⁻¹⁰g/cm², the deposition-preventing effect will not be sufficient, whilethe presence of more than 1×10⁻² g/cm² of the deposition-preventingagent will be liable to affect various test values.

The material for the vessel of the blood test ware of the invention canbe any of thermoplastic resint thermosetting resin, modified naturalresin, and glass. The thermoplastic resin mentioned above includes butis not limited to polyethylene, polypropylene poly(4-methylpentene-1),polystyrene, poly(methyl methacrylate), poly(vinyl chloride),poly(ethylene terephthalate), poly(butylene terephthalate),poly(styrene-co-acrylonitrile), poly(styrene-co-maleic anhydride),poly(styrene-co-acrylic acid), poly(styrene-co-methyl methacrylate),poly(ethylene-co-propylene), poly(ethylene-co-acrylic acid),poly(ethylene-co-acrylic ester), poly(vinyl acetal), and poly(vinylbutyral). The thermosetting resin mentioned above includes but is riotlimited to unsaturated polyester resin, epoxy resin, and epoxy-acrylateresin. The modified natural resin includes but is not limited tocellulose acetate, cellulose propionate, cellulose acetate butyrate,ethylcellulose, and ethylchitin.

The blood test ware of the present invention can be manufactured bycausing the blood component deposition-preventing agent of the inventionto be present on the inner wall of a blood test vessel or tube by avariety of alternative methods. Thus, for example, the method whichcomprises kneading the blood component deposition-preventing agent ofthe invention into a plastic batch for the molding of a vessel andmolding the kneaded mixture by the injection molding, blow molding orother technique and the method which comprises dissolving the bloodcomponent deposition-preventing agent in pure water or alcohol, applyingthe solution to the inside wall of the vessel by spray coating or dipcoating, and drying the coat can be mentioned.

In addition to the presence of the blood component deposition-preventingagent comprising the random copolymer according to the presentinvention, the blood test ware of the present invention may have aserum/plasma separator comprising a thixotropic fluid or a separatormaterial functioning as a partition between the serum or plasma layerand the blood solid component layer.

The serum/plasma separator includes a composition comprising liquidacrylic resin, chlorinated polybutene or liquid dicyclopentadiene (DCPD)as a matrix and a finely divided inorganic powder such as microfinesilica, alumina or glass particles as an auxiliary component forspecific gravity adjustment and thixotropy.

By allowing such a partition-forming agent to be present, the storagelife of serum or plasma can be increased without confounding testvalues.

The blood component deposition-preventing matrix of the presentinvention comprises a support and, as disposed on its surface, a bloodcomponent deposition-preventing agent comprising the random copolymeraccording to the first aspect of the present invention. The bloodcomponent deposition-preventing matrix is put to use as accommodated ina blood test tube or vessel. The support of said blood componentdeposition-preventing matrix can be any of the known supports. The shapeof the support is not limited and may, for example, be pellets, a sheet,a nonwoven fabric, or a woven fabric. The possible raw material of thesupport includes thermoplastic resin, thermosetting resin, and modifiednatural resin, among others. The thermoplastic resin mentioned justabove includes but is not limited to polyethylene, polypropylene,poly(4-methylpentene-1), polystyrene, poly(methyl methacrylate),poly(vinyl chloride), poly(ethylene terephthalate), poly(butyleneterephthalate), poly(styrene-co-acrylonitrile), poly(styrene-co-maleicanhydride), poly(styrene-co-acrylic acid), poly(styrene-co-methylmethacrylate), poly(ethylene-co-propylene), poly(ethylene-co-acrylicacid), poly(ethylene-co-acrylic enter), poly(vinyl acetal), andpoly(vinyl butyral). The thermosetting resin includes unsaturatedpolyester resin, epoxy resin, and epoxy-acrylate resin.

The amount of said blood component deposition-preventing agentcomprising the random copolymer on the surface of said blood componentdeposition-preventing support is preferably in the range of 1×10⁻¹⁰ to1×10² g/cm². If the amount of the blood component deposition-preventingagent is less than 1×10⁻¹⁰ g/cm², the deposition-preventing effect willnot be sufficient, while the presence of more than 1×10⁻² g/cm² of thedeposition-preventing agent will be liable to affect various testvalues.

The blood component deposition-preventing matrix of the presentinvention can be manufactured by causing the blood componentdeposition-preventing agent of the invention to be present on thesurface of a support by a variety of alternative methods. Thus, forexample, the method which comprises kneading the blood componentdeposition-preventing agent of the invention into a plastic batch forthe fabrication of the support and molding the kneaded mixture by theinjection molding, blow molding or other technique and the method whichcomprises dissolving the blood component deposition-preventing agent inpure water or alcohol, applying the solution to the surface of thesupport by spray coating or dip coating, and drying the coat can bementioned.

While the blood component deposition-preventing agent of the presentinvention can be used independently as described a above, it can be usedin combination with an adsorbent inorganic material, e.g. mineralsubstances such as glass, kaolin, bentonite, silica, cerite, etc., or incombination with an organic blood coagulation-accelerating substancesuch as ellagic acid. It can also be used in combination with ananticoagulant such as an ethylenediaminetetraacetic acid salt, citricacid salt, heparin salt, oxalic acid salt, or the like or anantiglycolytic agent such as fluorides, mannose, and so on.

When poly(vinylpyrrolidone-co-vinyl acetate) is selected as the bloodcomponent deposition-preventing agent of the invention and used incombination with said adsorbent inorganic material comprising at leastone member of the group consisting of glass, kaolin, bentonite, silicaand cerite, the vinylpyrrolidone content of saidpoly(vinylpyrrolidone-co-vinyl acetate) is preferably in the range of 10to 70 mol %. If the proportion of vinylpyrrolidone is less than 10 mol%, the agent will be stuck to the inside wall of the vessel so that noclot-exfoliating effect can be obtained. If the proportion ofvinylpyrrolidone exceeds 70 mol %, the agent will dissolve into theblood and not remain on the inner surface of the vessel so that noclot-exfoliating effect can be obtained.

The adsorbent inorganic material mentioned above is preferably amaterial not containing particles larger than 50 μm and having a meanparticle diameter of not more than 30 μm. Particularly for shorteningthe clotting time, the adsorbent inorganic substance is preferablysilica and a porous silica containing not less than 20 weight % of anamorphous fraction is particularly preferred. Such an adsorbentinorganic substance promotes activation of blood coagulation factors oncontact with blood and, also, accelerates aggregation of platelets.

In such cases, the amount of poly(vinylpyrrolidone-co-vinyl acetate) tobe present on the inside wall of the blood test vessel is preferably inthe range of 1×10⁻¹⁰ to 1×10⁻² g/cm². On the other hand, the amount ofthe adsorbent inorganic substance to be present on the inside wall ofthe blood test vessel is preferably in the range of 1×10⁻⁶ to 1×10⁻²g/cm². If it in less than 1×10⁻² g/cm², no bloodcoagulation-accelerating effect will be obtained. If the limit of 1×10⁻²g/cm² is exceeded, chances for inaccurate tests will be increased. Thecombined amount of said two materials is preferably not greater than1×10⁻² g/cm².

With the blood test ware carrying the poly(vinylpyrrolidone-co-vinylacetate) and adsorbent inorganic substance in combination, the bloodcoagulation factors are rapidly activated so that the clotting time isconsiderably shortened and, at the same time, the adhesion of theresulting clot to the inside wall of the blood test vessel issuccessfully prevented. As consequences, release of the serum from theclot is assisted, contamination of the serum with components of the clotis eliminated, and the serum yield is remarkably increased.

In order that said adsorbent inorganic substance may effectively exhibitits blood coagulation-accelerating action, each of the linseed oilabsorption value, BET specific surface area value, and resistivity valueis preferably within a certain range.

The linseed oil absorption value and BET specific surface area valuerepresent the magnitude of surface area of the adsorbent inorganicsubstance and the surface area value is also related with the degree ofsurface porosity of the adsorbent inorganic substance. Therefore, thedegree of surface porosity can be known from the oil absorption andspecific surface area values. The preferred adsorbent inorganicsubstance for use in the present invention preferably has a linseed oilabsorption value of 20 to 40 ml/100 g and a BET specific surface areavalue of 5000 to 30000 cm²/g.

The linseed oil absorption value is the value measured in accordancewith Japanese Industrial Standards (JIS) K-5101. The BET specificsurface area value means the value found by determining the amount ofgas which completely covers the surface as a monomolecular layer fromthe amount of gas adsorbed on the surface of an adsorbent inorganicsubstance, the prevailing equilibrium pressure, and the saturation vaporpressure of adsorbed gas and multiplying the result by the meansectional area of adsorbed gas molecules. As the adsorption gas,nitrogen gas, oxygen gas, argon gas, methane gas, etc. can beselectively employed. By this procedure, the Surface area inclusive offine pores which cannot be measured by the linseed oil absorption methodcan be determined. In the coagulation of blood, factor XII, that is tosay the contact factor, is activated but for this activation it isnecessary that the three substances of factor XII, prekalikrein andmacromolecular kininogen must form a complex and be adsorbed on thesurface of a foreign matter and it is said that the adsorption in thedeficiency in one or two of the three does not result in the activation.

In this connection, when an adsorbent inorganic substance used for thepurpose of accelerating blood coagulation is a substance having a varylarge surface area, the free factor XII, prekalikrein and macroolecularkininogen not forming a complex are adsorbed in an increased proportionon its surface, that is to say the proportion of the tripartite complexnecessary for the activation of factor XII is decreased so that theblood coagulation-accelerating effect is rather sacrificed. Converselywhen the surface area of the adsorbent inorganic substance is too small,the probability of adsorption of coagulation factors is decreased sothat the desired blood coagulation-accelerating effect cannot beexpected.

Therefore, the preferred adsorbent inorganic substance has a linseed oilabsorption value of 20 to 40 ml/100 g and a BET specific surface areavalue of 5000 to 30000 cm²/g.

The preferred resistivity value of the adsorbent inorganic substance isnot larger than 1×10¹⁰ Ω·cm and, for still better results, not largerthan 5×10⁴ Ω·cm. The resistivity value is the reciprocal of theelectrical conductivity value and represents the value at atmospherictemperature. It is supposed that the above resistivity of the adsorbentinorganic substance contributes to a sustained alignment of electricpotential distribution between the protein and the adsorbent inorganicsubstance and prevention of change in the conformation of the protein.

When poly(vinylpyrrolidone-co-vinyl acetate) as said blood componentdeposition-preventing agent and a salt of ethylenediaminetetraaceticacid, a salt of citric acid, a heparin salt, a salt of oxalic acid, orthe like as said blood anticoagulant are used in combination, the vinylacetate content of said poly(vinylpyrrolidone-co-vinyl acetate) ispreferably in the range of 30 to 90 mol %.

The ethylenediaminetetraacetate mentioned above can be any of thosesalts which are conventionally employed, such as disodiumethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate,tripotassium ethylenediaminetetraacetate, and so on.

The salt of citric acid can also be the salt conventionally used as ablood anticoagulant and may for example be trisodium citrate.

The heparin salt mentioned above may also be a salt conventionally usedas an Anticoagulant such as heparin sodium, heparin lithium, etc.

The salt of oxalic acid mentioned above can also be any salt that isconventionally used as an anticoagulant, thus including sodium oxalate,potassium oxalate, etc.

Any of the same known techniques as described hereinbefore can be usedfor causing said blood coagulation accelerator, blood anticoagulant, andantiglycolytic agent to be present on the inside wall of a blood testvessel or on the surface of a matrix to be accommodated in the bloodtest vessel. An exemplary procedure comprises causing the bloodcomponent deposition-preventing agent of the invention to be present onthe inside wall of a blood test vessel or the surface of a support inthe first place and, then, applying the blood coagulation accelerator,blood anticoagulant and/or antiglycolytic agent to the inside wall orsurface by way of spray-coating or dipping. An alternative procedurecomprises dissolving or suspending all the components in a suitablemedium, applying the solution or suspension to the substrate surface byway of spray-coating or dipping, and drying the coat.

Since the blood component deposition-preventing agent of the presentinvention is a random copolymer comprising 10 to 90 mol % of a monomercomponent (a) which would give a water-soluble homopolymer and 90 to 10mol % of a monomer component (b) which would give a water-insolublehomopolymer, it is structurally distinct from the known block copolymerof a hydrophilic monomer component such as a nonionic surfactant with ahydrophobic monomer component or the known graft polymer correspondingto such a block copolymer. While the nonionic surfactant mentioned aboveis coming into popular use as a useful sensitizer in immunoserologicaltests, the blood component deposition-preventing agent of the presentinvention is substantially free from the action of a sensitizer and, assuch, does not induce test errors such as false positive reactions.

In the second aspect of the present invention, a blood coagulationaccelerator is provided by supporting an antimicrobial metal on asupport or carrier and the resulting antimicrobial composition which issubstantially insoluble in blood is employed.

The support mentioned above must be eliminated from the serum atcentrifugation after the coagulation of blood for separation of serum.Since the specific gravity of human serum is 1.02 to 1.03, the abovesupport should have a specific gravity of not less than 1.03 andpreferably not less than 1.05.

The support material is not critical in kind only if the aboverequirement in regard to specific gravity is satisfied, thus including avariety of inorganic materials such as zeolite, montmorillonite,ceramics, glass, insoluble phosphates, etc. and a variety of organicmaterials such as graphite and ion exchange resins. Particularlypreferred are silicic acid compounds or silica series substances such aszeolite, montmorillonite, ceramics, etc. and insoluble phosphates, allof which per se have blood coagulation-accelerating properties as well.

The antimicrobial metal for use in the present invention is notparticularly limited, thus including the corresponding salts andorganometal compounds whose metal elements are copper and silver whichbelong to the Ib group of the periodic table of the elements, zinc,cadmium and mercury in the lib group, germanium, tin and lead in the IVagroup, lanthanids such as cerium and so on. In view of the balancebetween toxicity and utility, silver, copper, zinc, and cerium arepreferred.

The antimicrobial composition contained in the blood coagulationaccelerator according to the second aspect of the present inventioncomprises said support and, as supported thereby, said bacteriostaticmetal. The mode of supporting of said bacteriostatic metal on thesupport includes ion exchange, complex formation, and inclusion (as aclathlate). Other supporting modes are not satisfactory because of therisk of release of the metal into the blood.

The antimicrobial composition mentioned above is preferably in the formof a finely divided powder with a large surface area and the preferredparticle size is 0.01 to 500 μm . If the particle diameter is less than0.01 μm , a higher bacteriostatic action can be expected but in thecentrifugation step following completion of blood coagulation forseparation of serum, the composition may remain in serum under theroutine centrifugal conditions of about 1000 to 1800 G×5 minutes tocause clouding of the serum and other troubles such that the supportedmetal and the metal inherently contained in the serum are assayedtogether to introduce a positive error to the test value.

On the other hand, If the particle diameter exceeds 500 μm , theantimicrobial composition tends to be dispersed unevenly in thepreparation of a suspension of the blood coagulation accelerator, withthe result that the expression of antimicrobial activity is localizednear the surface of the antimicrobial composition ao that nobacteriostatic/fungistatic effect can be expected. Particularlypreferred is an antimicrobial composition having a mean particlediameter of 0.01 to 50 μm.

The minimum dose of said antimicrobial composition, like that ofantisepatic and antifungal agents in general, can be chosen according toMBC (minimal bactericidal concentration) and MFC (mininmal fungicidalconcentration). However, it is preferable to insure that theconcentration of the antimicrobial agent in a suspension of the bloodcoagulation accelerator of the invention in purified water, forinstance, will be not less than 0.1 μg/ml.

As to the maximum dose of said antimicrobial composition, objectionableevents such as hemolysis would be encountered if a large amount ofinsoluble matter finds its way into the blood. Therefore, it ispreferable to determine the formulation so that the concentration inblood in the event of release into the blood will not exceed 0.5 g/ml.

The above-mentioned blood coagulation accelerator can be prepared bymixing the above antimicrobial composition with a mineral bloodcoagulation-accelerating agent such as glass, kaolin, bentonite, silica,cerite, etc. or an organic blood coagulation-accelerating substance suchas ellagic acid.

The above blood coagulation accelerator can be suspended in pure wateror physiological saline to prepare a suspension and this suspension becontacted with the sample blood to shorten the clotting time.

Furthermore, the above blood coagulation accelerator can be used toconstruct a blood test ware having high blood coagulation-acceleratingactivity by suspending the accelerator in pure water or alcohol/purifiedwater and spraying the inside wall of the blood test vessel with thesuspension or immersing a support such as a nonwoven fabric or a plasticsheet in said suspension and, after drying and cutting the support tosize, accommodating the cutting in the blood test vessel.

The above-mentioned blood coagulation-accelerating suspension maycontain a water-soluble macromolecular compound such aspolyvinylpyrrolidone or modified cellulose as a binder for thecoagulation accelerator or a viscosity control agent for the suspension.

The antimicrobial composition comprising silver, copper, zinc or thelike as immobilized on zeolite, montmorillonite, ceramic, insolublephosphate or the like, which is contained in the blood coagulationaccelerator of the present invention destroys microorganisms invadingthe purified water or alcohol/purified water in which the acceleratorhas been suspended and is capable of preventing microbial contaminationduring storage of the blood test ware manufactured using the acceleratorand, yet, will no dissolve into the blood so that the function of theblood coagulation accelerator is not adversely affected, nor does itinterfere with blood examination values. Moreover, because theantimicrobial composition itself has blood coagulation-acceleratingactivity, the specific activity of the whole blood coagulationaccelerator as a complex artifact is not compromised. The reason whysufficient antimicrobial efficacy can be expected despite the fact thatthe antimicrobial metal contained in the antimicrobial composition ofthe invention is little released in the form of free ions is probablythat active oxygen is generated in the vicinity of the supported metal.

The essential feature of the third aspect of the present inventionresides in the following constructions of (3-1), (3-2), (3-3), (3-4),and (3-5).

(3-1)

A blood test ware characterized in that 1×10⁻¹⁰ to 1×10⁻² g/cm² of apolyvinylpyrrolidone having a weight average molecular weight of 100000to 2000000 is disposed on the inside wall of a plastic vessel and, inaddition, at least one blood anticoagulant selected from the groupconsisting of the salts of ethylenediaminetetraacetic acid, heparin,citric acid, and oxalic acid in disposed in said plastic vessel.

(3-2)

A blood test ware characterized in that a composition comprising thefollowing components (1), (2), and (3) is disposed on the inside wallthereof, and a blood test matrix characterized in that a compositioncomprising the following components (1), (2), and (3) is disposed on thesurface thereof, and which is substantially insoluble in blood andphysicochemically substantially inert to blood and has a specificgravity of not less than 1.03 and a maximum projected length of not lessthan 1 mm.

(1) a polyvinylpyrrolidone having a weight average molecular weight of100000 to 2000000

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter of 1 mμ to 100 μ

(3-3)

A blood test ware characterized in that a composition comprising thefollowing components (1), (2), and (3) is disposed on the inside wallthereof, and a blood test matrix characterized in that a compositioncomprising the following components (1), (2), and (3) is disposed on thesurface thereof, and which is substantially insoluble in blood andphysicochemically substantially inert to blood and has a specificgravity of not less than 1.03 and a maximum projected length of not lessthan 1 mm.

(1) a random copolymer comprising 10 to 90 mol % of a monomer component(a) the homopolymer of which is water-soluble and 90 to 10 mol % of amonomer component (b) the homopolymer of which is water-insoluble

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter of 1 mμ to 100 μ

(3-4)

A blood test ware characterized in that a composition comprising thefollowing components (1), (2), and (3) is disposed on the inside wallthereof, and a blood test matrix characterized in that a compositioncomprising the following components (1), (2), and (3) is disposed on thesurface thereof, and which is substantially insoluble in blood andphysicochemically substantially inert to blood and has a specificgravity of not less than 1.03 and a maximum projected length of not lessthan 1 mm.

(1) a nonionic surfactant

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter of 1 mμ to 100 μ

(3-5)

A blood test ware characterized in that a composition comprising thefollowing components (1), (2), (3), and (4) is disposed on the insidewall thereof, and a blood test matrix characterized in that acomposition comprising the following components (1),(2), (3), and (4) isdisposed on the surface thereof, and which is substantially insoluble inblood and physicochemically substantially inert to blood and has aspecific gravity of not less than 1.03 and a maximum projected length ofnot less than 1 mm.

(1) at least one blood component deposition-preventing agent selectedfrom the group consisting of silicone oil, polar group-containingmodified silicone oil, polyhydric alcohol partial esters, polyhydricalcohol complete esters, and poly(propylene oxide)

(2) a water-soluble macromolecular compound

(3) a blood anticoagulant

(4) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has. a specificgravity of not less than 1.08 and a particle diameter of 1 mμ to 100 μ

The third aspect of the present invention is now described in detail.

The blood test ware (3-1) comprises a plastic vessel and, as disposed onthe inside wall thereof, 1×10⁻¹⁰ to 1×10⁻² g/cm² of apolyvinylpyrroidone having a weight average molecular weight of 100000to 2000000. If the amount of said polyvinylpyrrolidone Is less than1×10⁻¹⁰ g/cm², it will be impossible to obtain the desired bloodcomponent deposition-preventing effect, while more than 1×10⁻² g/cm² ofpolyvinylpyrrolidone will interfere with blood examinations.

The weight average molecular weight of polyvinylpyrrolidone can bedetermined by the conventional methods such as ultracentrifugation orthe light scattering method. As an alternative, the viscosity averagemolecular weight (Mv) is first calculated from the viscosity value knownas K value by means of the following equation (1) and the weight averagemolecular weight (Mw) is then calculated by means of the followingequation (2) [V. Buehler, U. Klodwig. Acta Pharm., Techn., 30. No, 4(1984)].

Mv=22.22×(K+0.075×K²)^(1.65)  (1)

Mw≈Mv  (2)

The weight average molecular weight of said polyvinylpyrrolidone is100000 to 2000000. If it is less than 100000, the polyvinylpyrrolidonewill dissolve into blood and disappears from the inside wall of theblood test ware so that the blood component deposition-preventing effectcannot be obtained, if the molecular weight exceeds 2000000, spray orother coating workability is sacrificed. Therefore, the above range isessential. The preferred range is 300000 to 1500000 and. the still morepreferred range is 600000 to 1500000.

Further disposed in this blood test ware is at least one bloodanticoagulant selected from the group consisting of the salts ofethylenediamninetetraacetic acid, heparin, citric acid, and oxalic acid,and fluorides.

The above-mentioned salt of ethylenediaminetetraacetic acid can be anyof the salts which are Conventionally used as blood anticoagulants, suchas disodium ethylenediaminetetraacetate, dipotassiumethylenediaminetetraacetate, and tripotassiumnethylenediaminetetraacetate, among others.

The above-mentioned salt of citric acid can also be any of the saltsconventionally used as blood anticoagulants, such as trisodium citrate,among others.

The heparin salt mentioned above can be a salt of heparin which iscommonly used as a blood anticoagulant, thus including heparin sodium,heparin lithium and so on.

The above-mentioned salt of oxalic acid can be any of those oxalateswhich are conventionally used as blood anticoagulants, thus includingsodium oxalate and potassium oxalate, among others.

Among the fluorides mentioned above are sodium fluoride and potassiumfluoride which are conventionally used as antiglycolytic agents.

In the above blood test ware may be further accommodated a materialcapable of establishing a partition between a plasma layer and a solidcomponent layer, such as a plasma separator comprising a thixotropicfluid or a separating member. The plasma separator may for example be acomposition comprising chlorinated polybutene or dicyclopentadiene(DCPD) resin as a main component and a finely divided inorganic powdersuch as powdered silica, alumina or glass as a viscosity control andthixotropic agent. When such a partitioning material is provided, theblood test were can store the plasma for a long time without adverseeffects on test data.

The method of using the blood test ware comprises drawing the bloodsample into the ware and after thorough mixing of the anticoagulant andthe blood, centrifuging the ware to separate the plasma.

Since a specified quantity of a specified grade of polyvinylpyrrolidoneis present on the inside wall of the vessel in the above blood testware, corpuscular elements and proteins in the blood are prevented fromadhering to the inside wall surface. Moreover, since the bloodanticoagulant is accommodated in the vessel, coagulation of the blood isprevented. Furthermore, in the embodiment where a partition-formingsubstance is provided within the vessel, the plasma can be stored instable condition for a long time without influences on test values.

The blood test ware (3-2) comprises a vessel and, as disposed on theinside wall of said vessel, a composition comprising the followingcomponents (1), (2) and (3), and the corresponding blood test matrixcomprises a support and, as disposed on its surface, a compositioncomprising the following components (1), (2) and (3), and issubstantially insoluble in blood and physicochemically substantiallyinert to blood and has a specific gravity of not less than 1.03 and amaximum projected length of not less than 1 mm.

(1) a polyvinylpyrrolidone having a weight average molecular weight of100000 to 2000000

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle size within the range of 1mμ to 100 μ

The description for (3-1) applies to said polyvinylpyrrolidone having aweight average molecular weight of 100000 to 2000000 (1).

The description for (3-1) applies to said blood anticoagulant (2).

The finely divided powder (3) is now described. using a medium which isa good solvent for both the polyvinylpyrrolidone and bloodanticoagulant, a homogeneous solution can be prepared. However, when theinside wall of a plastic vessel is coated with such a solution byspray-coating or dip-coating and the coated vessel is allowed to standin the upright position, the solution Is not retained on the inside wallbut flows down to the bottom of the vessel. In that event, the solutionforms a thick dry film on the bottom to seriously interfere withredissolution of the anticoagulant in the blood so that the bloodundergoes local coagulation and in the subsequent step of accommodatinga plasma separator most of the anticoagulant is buried under the plasmaseparator and fails to contact the blood introduced so that the expectedeffect is not accomplished. The finely divided powder (3) has theproperty to considerably improve the retention of the dispersion on theplastic vessel by suppressing the sagging tendency so that the aboveproblem is neatly solved.

The above effect of the finely divided powder (3) is probably attributedto the following. Thus, as the finely divided powder is adsorbed on theinside wall surface, a large number of fine projections and recesses areformed on the inside wall to increase the surface area and theretentivity of the solution is increased as the result of surfacetension.

If the specific gravity of said finely divided powder (3) is less than1.08, the powder (3) could remain in the plasma even after centrifugalseparation of the blood to interfere with blood tests. Therefore, thespecific gravity is restricted to 1.08 or more.

If the particle diameter of finely divided powder (3) is less than 1 mμthe fine particles tend to form a dense film upon concentration todryness to interfere with redissolution of the blood anticoagulant. Onthe other hand, if 100 μ is exceeded, the separation and sedimentationrate of particles in a mixed dispersion of polyvinylpyrrolidone andblood anticoagulant is increased to sacrifice the retentivity on theinside wall surface of the vessel. Therefore, the particle diametershould be limited to the above range. The preferred range is 1 to 50 μ.

The level of addition of finely divided powder (3) is not particularlycritical but a sufficient effect can be obtained when it is present in aproportion of 5 weight % or less.

The material of said finely divided powder (3) is not so critical andcan be any of such materials as, for example, poly(meth)acrylic acidesters, poly(vinyl chloride), fluororesins, polyamides, polyesters,polyoxyalkylenes, polyurethane, urea resin, melamine resin, epoxy resin,phenolic resin, cellulose, chitin, modified cellulose, modified chitin,and their copolymers and crosslinked polymers. Even polystyrene,polyethylene and polypropylene which cannot be used alone on account oftheir low specific gravities can be utilized when a conventionalinorganic filler such as silica, talc or the like has been kneaded intothem for specific gravity adjustment. These powders can be manufacturedin the conventional manner, for example by suspension polymerization orpulverization and size selection. In the above blood test ware may bedisposed a material capable of providing a partitioning wall between theplasma layer and the solid component layer, such as a plasma separatorcomprising a thixotropic fluid or a separatory member.

The blood test matrix according to (3-2) carries a compositioncomprising the above-mentioned components (1), (2) and (3) on itssurface and provides for the same effect as the blood test waredescribed above. The blood test matrix should not affect the bloodexamination and, therefore, is designed to be substantially insoluble inblood and physicochemically substantially inert to blood. This bloodtest matrix has a specific gravity of not less than 1.03 and a maximumprojected length. of not less than 1 mm. If the specific gravity is lessthan 1.03, the matrix floats without sinking in the blood sample tointerfere with the examination. If the maximum projected length is lessthan 1 mm, workability is sacrificed.

The above blood test matrix is used as accommodated in the blood testvessel. The known support materials can be used for the fibrication ofsaid blood test matrix. There is no particular limitation on matrixconfiguration, and pellets, sheet, nonwoven cloth, woven cloth, etc. canbe mentioned as examples. The raw material is not particularlyrestricted, either, and a variety of materials similar to thosementioned for said component (3) can be utilized.

The blood test ware according to (3-3) comprises a vessel and, asdisposed on its inside wall surface, a composition comprising thefollowing components (1), (2) and (3). The corresponding blood testmatrix comprises a support and, as disposed on its surface, acomposition comprising the following components (1), (2) and (3), and issubstantially insoluble in blood and physicochemically inert to bloodand has a specific gravity of not less than 1.03 and a maximum projectedlength of not less than 1 mm.

(1) a random copolymer comprising 10 to 90 mol % of a monomer components(a) which would give a water-soluble homopolymer and 90 to 10 mol % of amonomer component (b) which would give a water-insoluble homopolymer.

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter in the range of 1mμ to 100 μ

The above-mentioned random copolymer (1) comprises a monomer component(a) which would give a water-soluble homopolymer and a monomer component(b) which would give a water-insoluble homopolymer. The monomercomponent (a) which would give a water-soluble homopolymer that can beused includes but is not limited to vinylpyrrolidone, vinyl alcohol,ethylene oxide, salts of acrylic acid, salts of styrenesulfonic acid,salts of vinylphosphonic acid, s allylamine salts, hydroxymethyl(meth)acrylate, glycosylethyl (meth)acrylate, saccharides such asglucose, and amino acids such as glutamic acid. These monomers can beUsed alone or as a mixture.

The monomer component (b) which would give a water-insoluble homopolymerthat can be used includes but is not limited to ethylene, propylene,propyleneoxide, vinyl acetate, vinyl chloride, alkyl (meth)acrylates,styrene, acrylonitrile, and acrolein. These monomers can be used aloneor as a mixture.

The random copolymer comprising said monomer components (a) and (b) canbe produced typically by the known addition polymerization reaction orpolycondensation reaction. In view of the availability of materials, itis advantageous to synthesize the random copolymer by usingvinylpyrrolidone or vinyl alcohol as the monomer component (a) and vinylacetate as the monomer component (b). As commercial products, LuviskolVA grade numbers VA73, VA64, VA55, VA37, and VA28 are available fromBASF as typical vinylpyrrolidone-vinyl acetate random copolymers andUnitika Poval grade numbers n-180, UMR-10M, UMR-30L, and UMR-150L areavailable from Unitika Ltd. as typical vinyl alcohol-vinyl acetaterandom copolymers.

In the above random copolymer, the proportion of monomer component (a)the homopolymer of which is water-soluble is within the range of 10 to90 mol % and that of monomer component (b) the homopolymer of which iswater-insoluble is within the range of 90 to 10 mol %.

If the proportion of monomer component (a) the homopolymer of which iswater-soluble exceeds 90 mol %, the resulting random copolymer will notbe much different in characteristics from the homopolymer of monomercomponent (a) so that the rate of adsorption on the inside wall of theblood test ware and on the matrix surface is decreased and thesolubility in blood is too high. As a consequence, when blood is drawninto the blood test ware, the random copolymer is washed out from theinside wall of the ware or the matrix surface so that it cannot play therole of preventing deposition of blood components.

On the other hand, if the proportion of monomer component (a) whosehomopolymer is water-soluble is less than 10 mol %, the copolymer willnot be much different in characteristics from the homopolymer of monomercomponent (b) and be substantially insoluble in blood so that it may notplay the role of preventing deposition of blood components. When acombination of. this blood component deposition-preventing agent with ablood coagulation accelerator or a blood anticoagulant is applied to theinside wall of a blood test vessel or the matrix surface and dried, ablood-insoluble film is formed on the surface of the blood coagulationaccelerator or blood anticoagulant so that blood coagulation factorsXII, XI, etc. cannot bind to the surface of the blood coagulationaccelerator, with the result that the coagulation of blood is nothastened and the reduced solubility of the blood anticoagulant leads toan insufficient anticoagulant effect.

For the above reasons, the random copolymer for use in the presentinvention should be such that said monomer component (a) whosehomopolymer is water-soluble and said monomer component (b) whosehomopolymer is water-insoluble account for 10 to 90 mol % and 90 to 10mol % respectively.

The amount of said random copolymer is preferably 1×10⁻¹⁰ to 1×10⁻²g/cm². If it is less than 1×10⁻¹⁰ g/cm², deposition of corpuscularcomponents and proteins will not be sufficiently precluded. If theamount of the random copolymer exceeds 1×10⁻² g/cm², various testparameter values will be confounded.

The above blood test ware according to (3-4) comprises a vessel and, asdisposed on its inside wall, a composition comprising the followingcomponents (1), (2) and (3) and the corresponding blood test matrixcomprises a support and, as disposed on its surface, a composition ofthe following components (1), (2) and (3) and is substantially insolublein blood and physicochemically substantially inert to blood and has aspecific gravity of not less than 1.03 and a maximum projected length ofnot less than 1 mm.

(1) a nonionic surfactant

(2) a blood anticoagulant

(3) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter in the range of 1mμ to 100 μ

The above nonionic surfactant (1) that can be used includes but is notlimited to ethylene glycol/propylene glycol series, alkyllalkylene oxideseries, and alkylene oxide/silicone series block copolymers and graftcopolymers, inclusive of the corresponding modified polymers.Particularly preferred are surfactants with HLB (hydrophilic-lypophilicbalance) numbers not less than 10.

The preferred proportion of said nonionic surfactant (1) is 1×10⁻¹⁰ to1×10⁻² g/cm². If it is less than 1×10⁻¹⁰ g/cm², deposition ofcorpuscular components and proteins will not be sufficiently precluded.If the amount of the surfactant exceeds 1×10⁻² g/cm², various testparameter values could be confounded.

The blood test ware according to (3-5) comprises a vessel and, asdisposed on the inside wall surface thereof, a composition comprisingthe following components (1), (2), (3) and (4), and the correspondingblood test matrix comprises a support and, as disposed on its surface, acomposition comprising the following components (1), (2), (3) and (4)and is substantially insoluble in blood and physiochemicallysubstantially inert to blood and has a specific gravity of not less than1.03 and a maximum projected length of not less than 1 mm.

(1) At least one blood component deposition-preventing agent selectedfrom the group consisting of silicone oil, modified silicone oilcontaining polar groups, partial esters of polyhydric alcohols, completeesters of polyhydric alcohols, and poly(propylene oxide)

(2) a water-soluble macromolecular substance

(3) a blood anticoagulant

(4) a finely divided powder which is substantially insoluble in bloodand physicochemically substantially inert to blood and has a specificgravity of not less than 1.08 and a particle diameter In the range of 1mμ to 100 μ

The silicone oil (1) that can be used includes but is not limited todimethylpolysiloxane, methylhydrogenpolysiloxane, andmethylphenylpolyslloxane. The polar group-modified silicone oil (1)includes oils obtainable by introducing polar groups such as hydroxyl,amino, carboxyl, epoxy, etc. into dimethylpolysiloxane,methylhydrogenpolysiloxane, methylphenylpolysiloxane and other siliconeoils.

The above-mentioned partial or complete esters of polyhydric alcohols(1) are compounds available on introduction of fatty acid molecules tosome or all of the alcoholic hydroxyl functions of the respectivepolyols such as glycerol, sorbitol, polyphenol, etc.

The blood component deposition-preventing agent (1) includes, inaddition to the substances mentioned above, poly(propylene oxide) andother substances.

The preferred amount of said blood component deposition-preventing agent(1) is 1×10⁻¹⁰ to 1×10⁻² g/cm². If the amount of (1) is less than1×10⁻¹⁰ g/cm², no sufficient deposition-preventing effect on corpuscularelements and proteins will be obtained. If it exceeds 1×10⁻² g/cm²,various test parameter values will be interfered with.

The water-soluble macromolecular compound (2) that can be used includesbut is not limited to poly(ethylene oxide), poly(vinyl alcohol),polyvinylpyrrolidone, poly(sodium acrylate), polyethyleneimine, sodiumalginate, starch, pullulan, methylcellulose, hydroxyethylcellulose,hydroxypropylcellulose, carboxymethylcellulose, cellulose acetatephthalate, gum arabic, guns tragacanth, locust bean gum, guar gum,pectin, carrageenan, phaseleran, tamarind seed polysaccharide, glue,gelatin and casein. Particularly preferred are polyvinylpyrrolidone,poly(ethylene oxide) and poly(vinyl alcohol).

The water-soluble macromolecular compound (2) serves to prevent thewater-insoluble matter from covering the blood anticoagulant to inhibitits solubility in blood.

The preferred amount of said water-soluble macromolecular compound (2)is 1×10⁻¹⁰ to 1×10⁻² g/cm². If it is less than 1×10⁻¹⁰ g/cm², nosufficient deposition-preventing effect on corpuscular elements andproteins will be obtained.

If the amount of the compound exceeds 1×10⁻² g/cm², various testparameter values could be confounded.

EXAMPLES

The following examples are intended to describe the present invention Infurther detail and should by no means be construed as defining the scopeof the invention.

Examples 1-1 to 1-27

The present invention is now described with reference to the examplesusing the 9 kinds of random copolymers listed in Table 1-1.

TABLE 1-1 Copolymer Manu- Trade Grade Mol % of structure facturer namedesignation monomer (b) Vinylpyrrolidone- BASF Luviskol VA73 35 vinylacetate VA VA64 46 random copolymer VA55 56 VA37 75 VA28 84 Vinylalcohol- UNITIKA Unitika E-180 10 vinyl acetate Poval UMR-10M 35 randomcopolymer UMR-30L 60 UMR-150L 80

For use as the blood component deposition-preventing agent, aqueoussolutions of the vinylpyrrolidone-vinyl acetate random copolymers shownin Table 1-2 (only the grade numbers are shown in Table 1-2) in purewater were prepared to the concentrations (weight %) specified in Table1-2. Moreover, aqueous solutions of the vinyl alcohol-vinyl acetaterandom copolymers shown in Table1-3 (only the grade numbers are given inTable 1-3) in pure water were prepared to the concentrations (weight %)specified in Table 1-3. However, UMR-150L only was dissolved inmethanol.

About 50 μl of each solution was sprayed into a poly(ethyleneterephthalate) blood sampling tube of 10 ml capacity according to Table1-2 or Table 1-3 and dried in an air-current dryer at 60° C. to providea blood test ware. Fresh rabbit blood, 3 ml, was taken in each bloodtest ware and allowed to stand at the room temperature of 23 to 25° C.After complete coagulation of blood was confirmed at 4 hours, theadhesion of the clot to the inside wall of the blood test ware wasvisually evaluated. As a result, none of the tubes showed adhesion ofthe clot. Then, the tubes were centrifuged at 1300 G (25° C.) for 5minutes and the adhesion of the clot on the inside wall of the bloodtest ware was visually evaluated again. At the same time, the yield ofserum that separated out was determined. As a result, none of the tubesshowed adhesion of the clot to the inside wall. The serum yields areshown in Table 1-2 and Table 1-3.

TABLE 1-2 Serum yield Copolymer after Example Grade Concentration (%)centrifugation (ml) 1-1 VA73 0.05 1.4 1-2 VA73 1.00 1.4 1-3 VA73 5.001.5 1-4 VA64 0.05 1.4 1-5 VA64 1.00 1.5 1-6 VA64 5.00 1.5 1-7 VA55 0.051.5 1-8 VA55 1.00 1.5 1-9 VA55 5.00 1.4  1-10 VA37 0.05 1.5  1-11 VA371.00 1.4  1-12 VA37 5.00 1.4  1-13 VA28 0.05 1.4  1-14 VA28 1.00 1.4 1-15 VA28 5.00 1.4

TABLE 1-3 Serum yield Copolymer after Example Grade Concentration (%)centrifugation (ml) 1-16 E-180 0.05 1.4 1-17 E-180 1.00 1.4 1-18 E-1805.00 1.5 1-19 UMR-10M 0.05 1.4 1-20 UMR-10M 1.00 1.5 1-21 UMR-10M 5.001.4 1-22 UMR-30L 0.05 1.5 1-23 UMR-30L 1.00 1.5 1-24 UMR-30L 5.00 1.41-25 UMR-150L 0.05 1.4 1-26 UMR-150L 1.00 1.5 1-27 UMR-150L 5.00 1.4

Comparative Example 1-1

As the blood component deposition-preventing agent, a 0.05 weight %aqueous solution of vinylpyrrolidone homopolymer (manufactured by BASF,Luviskol K-30TM, weight average molecular weight 30000) was used in lieuof a 0.05 weight % solution of vinylpyrrolidone-vinyl acetate randomcopolymer. The test was carried out in otherwise the same manner as inExample 1-1. Four hours after blood sampling, complete coagulation ofblood was confirmed and the adhesion of the clot to the inside wall ofthe blood teat ware was visually evaluated. As a result, clot depositswere found. In the evaluation of clot deposits on the inside wall of theware after centrifugation, not only clot deposits were found but alsomarked hemolysis was observed. The serum yield after centrifugation was1.4 ml.

Comparative Example 1-2

As the blood component deposition-preventing agent, a 0.05 weight %aqueous solution of vinyl alcohol homopolymer (manufactured by UnitikaLtd., Unitika Poval (tradename) UF200G) was used in lieu of a 0.05weight % solution of vinylpyrrolidone-vinyl acetate random copolymer.The test was carried out in otherwise the same manner as in Example 1-1.Four hours after blood sampling, complete coagulation of blood wasconfirmed and the adhesion of the clot to the inside wall of the bloodtest ware was visually evaluated. As a result, clot deposits were found.In the evaluation of clot deposits on the inside wall of the ware aftercentrifugation, not only clot adhesion spots but also moderate hemolysiswas observed. The serum yield after centrifugation was 1.4 ml.

Comparative Example 1-3

As the blood component deposition-preventing agent, a 0.05 weight %methanolic solution of vinyl acetate-ethylene copolymer (manufactured byHoechst Gosei.Co.; mobinil (tradename) E45) was used in lieu of a 0.05weight % solution of vinylpyrrolidone-vinyl acetate random copolymer.The test was carried out in otherwise the same manner as in Example 1-1.Four hours after blood sampling, complete coagulation of blood wasconfirmed and the degree of adhesion of the clot to the inside wall ofthe blood test ware was visually evaluated. AS a result, marked adhesionwas found. In the examination of clot adhesion to the inside wall of theware after centrifugation, marked adhesion was found. The serum yieldafter centrifugation was 0.0 ml.

Comparative Example 1-4

The test procedure of Example 1-1 was repeated except that the warecarrying no blood component deposition-preventing agent (that is anuntreated poly(ethylene terephthalate) tube of 10 ml capacity) was usedas the blood test ware, Four hours after blood sampling, completecoagulation of blood was confirmed and the degree of adhesion of theclot to the inside wall of the blood test ware was visually evaluated.As a result, marked deposits were found. In the examination of clotadhesion to the inside wall of the ware after centrifugation, not onlymarked deposits of the clot but also hemolysis was observed. The serumyield after centrifugation was not more than 0.5 ml.

The results of Examples 1-1 to 1-27 and Comparative Examples 1-1 to 1-4indicate that whereas the poly(ethylene terephthalate) blood samplingtube (Comparative Example 1-4) inherently has the property to stronglybind blood components, the blood component deposition-preventing agentrandom copolymer of the present invention exhibits an excellentdeposition-preventing effect. The serum yields in Examples 1-1 to 1-27were approximately 50% of the whole blood, indicating that substantiallythe whole amount of serum was recovered. Comparative Examples 1-1 and1-2 correspond to the case where, of the monomer composition of therandom copolymer for use in the present invention, the proportion of themonomer component which would yield a water-insoluble homopolymer is 0mol %, and Comparative Example 1-3 correspond to the case in which theproportion of the same monomer component is 100 mol %. comparativeExamples 1-1 and 1-2 were obviously inferior giving punctuate blood clotdeposits on the inside wall, although the serum yield was not soaffected, and, in addition, showed hemolysis. In Comparative Example1-3, marked clot deposits were found and the recovery of serum wasinfeasible. All of the cases are deviating from the requirement of theinvention that the proportion of the monomer compound giving awater-insoluble homopolymer should be 10 to 90 mol %, thus failing toinhibit deposition of blood components.

Example 2-1

Using a vinylpyrrolidone-vinyl acetate random copolymer (manufactured byBASF, Luviskol (tradename) VA73, vinyl acetate content ca 36 mol %) asthe blood component deposition-preventing agent and a finely dividedsilica powder (mean particle diameter 4.0 μm, the linseed oil absorptiondetermined in accordance to JIS K 5101=30 ml/100 g, BET specific surfacearea 12000 cm²/g, resistivity as the reciprocal of electricalconductivity=2.6×10⁻⁴ Ω19 cm) as the blood coagulation-acceleratinginorganic adsorbent, a methanolic dispersion was prepared to therespective concentrations of 0.1 weight % and 1.0 weight %. Thisdispersion was sprayed onto the inside wall of a 10 ml polypropyleneblood sampling tube and dried to provide a blood test ware.

The amounts of deposition of the respective components per unit area ofthe inside wall surface of the tube were 2×10⁻⁶ g/cm² forvinylpyrrolidone-vinyl acetate copolymer and 2×10⁻⁵ g/cm² for finelydivided silica.

Fresh human blood, 8 ml, was taken into the above blood test ware andallowed to stand at 20° C. The time till complete loss of fluidity ofthe blood was measured as blood coagulation time for assessment ofclotting performance. After confirmation of coagulation, the sample wasimmediately centrifuged at 3000 rpm for 5 minutes and the serumseparability was evaluated. At the same time, the supernatant waspipetted to find the serum yield. The results are shown in Table 1-4.

Example 2-2

Using the vinylpyrrolidone-vinyl acetate random copolymer (manufacturedby BASF, Luviskol (tradename) VA28, vinyl acetate content ca 84 mol %)as the blood component deposition-preventing agent and the same finelydivided silica as used in Example 2-1 as the blood coagulationaccelerator, a methanolic dispersion was prepared to the respectiveconcentrations of 0.02 weight % and 1.0 weight %. This dispersion wassprayed onto the inside wall of a 10 ml polypropylene blood samplingtube and dried to provide a blood test ware.

The amounts of deposition of the respective components per unit area ofthe inside wall surface of the tube were 5×10⁻⁷ g/cm² forvinylpyrrolidone-vinyl acetate copolymer and 3×10⁻⁵ g/cm² for finelydivided silica.

Then, as in Example 2-1, clottability, serum separability, and serumyield were evaluated. The results are shown in Table 1-4.

Comparative Example 2-1

Using a vinylpyrrolidone homopolymer (manufactured by BASF,Luviskol(tradename) K30, weight average molecular weight 30000) as theblood component deposition-preventing agent and the same finely dividedsilica as used in Example 2-1 as tile blood coagulation accelerator, amethaniolic dispersion was prepared to the respective concentrations of0.1 weight % and 1.0 weight %. This dispersion was spray-coated on theinside wall of a 10 ml polypropylene. blood sampling tube and air-driedto provide a blood test ware.

The amounts of deposition of the respective components per unit area ofthe inside wall surface of the tube were 3×10⁻⁶ g/cm² forvinylpyrrolidone homopolymer and 3×10⁻⁵ g/cm² for finely divided silica.Then, as in Example 2-1, clottability, serum separability, and serumyield were evaluated. The results are shown in Table 1-4.

Comparative Example 2-2

Using the same finely divided silica as used in Example 2-1 as the bloodcoagulation accelerator but not using a blood componentdeposition-preventing agent, a methanolic dispersion containing 1.0weight % of silica was prepared. This dispersion was spray-coated on theinside wall of a 10 ml polypropylene blood sampling tube and air-driedto provide a blood test ware.

The amount of deposition of the above component per unit area of theinside wall surface of the ware was 3×10⁻⁵ g/cm².

Then, as in Example 2-1, clottability, serum separability, and serumyield were evaluated. The results are shown in Table 1-4.

TABLE 1-4 Blood coagulation Serum Serum yield time (min.) separability(ml) Example 2-1 20 good 4.2 Example 2-2 25 good 4.2 Comparative 35 poorNot recovered Example 2-1 Comparative 40 poor Not recovered Example 2-2

Example 2-1

Using the vinylpyrrolidone-vinyl acetate random copolymer (manufacturedby BASF, Luviskol (tradename) VA64, vinyl acetate content ca 46 mol %)as the blood component deposition-preventing agent, an aqueous solutionof 1.0 weight % concentration in pure water was prepared. On the otherhand, using the same finely divided silica as used in Example 2-1 as theblood coagulation accelerator, an aqueous dispersion of 1.0 weight tconcentration in pure water was prepared. About 50 μl of the aqueoussolution of vinylpyrrolidone-vinyl acetate random copolymer wasspray-coated on the inside wall of a polyethylene terephthalate) bloodsampling tube of 10 ml capacity and dried in an air-current dryer at 60°C. Then, about 50 μl of the aqueous suspension of finely divided silicawas further spray-coated and dried in an air-current dryer at 60° C. toprovide a blood test ware. Fresh rabbit blood, 5 ml, was taken in thisblood test ware and allowed to stand at the room temperature of 23 to25° C. The time till loss of fluidity of the blood and start ofseparation of serum was measured as blood coagulation time. Then, at 1hour after blood sampling and following centrifugation, the degree ofadhesion of the blood clot to the inside wall of the blood test ware wasvisually evaluated and the serum yield determined as in Example 1-1. Theresults are shown in Table 1-5.

Then, to confirm the influence of the blood componentdeposition-preventing agent on immunoserological parameter values, thewhole amount of the serum was transferred to a clean glass test tubeimmediately after completion of the above evaluation, Then, using HBs-Abtest reagent (manufactured by To a Medical Electronics Co., Ltd.) andFree T4 test reagent (manufactured by Kodak), HBs-Ab and Free T4 testswere performed on the serum for immunoserological assessment. The testresults were not false positive but negative as shown in Table 1-5.

Example 3-2

Except that a 1.0 weight % aqueous solution of vinyl alcohol-acetaterandom copolymer (manufactured by Unitika Ltd., Unitika Poval(tradename) UMR-30L, vinyl acetate content ca 60 mol %) in pure waterwas used in lieu of a 1.0 weight % solution of vinylpyrrolidone-vinylacetate random copolymer in pure water, a blood test ware was fabricatedin otherwise the sate manner as in Example 3-1 and the tests describedin Example 3-1 were carried out. The results are shown in Table 1-5.

Comparative Example 3-1

Except that the use of vinylpyrrolidone-vinyl acetate random copolymerwas omitted, a blood test ware was fabricated in otherwise the samemanner as in Example 3-1 (silica only was used), and the tests describedin Example 3-1 were carried out. The results are shown in Table 1-5.

However, immunoserological tests could not be performed because theblood clot was not separable from the inside wall of the blood samplingtube so that the serum could not be recovered.

Comparative Example 3-2

Except that a plain hard glass blood sampling tube of 10 ml capacity inlieu of the blood test ware comprising a polyethylene terephthalate)blood sampling tube containing said vinylpyrrolidone-vinyl acetaterandom copolymer and silica, the same tests as described in Example 3-1were performed. The results are shown in Table 1-5.

Comparative Example 3-3

Except that a 1.0 weight t solution of a polyether-modified silicone oilnonionic surfactant (manufactured bay Toray-Dow Corning Silicone,SH3749) in pure water in lieu of a solution of vinylpyrrolidone-vinylacetate random copolymer in pure water, the procedure of Example 3-1 wasotherwise repeated to provide a blood test ware and the tests describedin Example 3-1 were carried out. The results are shown in Table 1-5.

TABLE 1-5 Example Comparative Example 3-1 3-2 3-1 3-2 3-3 Bloodcoagulation 20 20 20 30 25 time (min.) Adhesion of clots at None NoneSignificant None None 1 hr after blood adhesion sampling Adhesion ofclots after None None Significant None None centrifugation adhesionSerum yield after 2.6 2.5 0.0 2.5 2.5 centrifugation (ml) HBs-Ab Neg-Neg- No recovery Negative False ative ative of serum; positive Free T4Neg- Neg- not tested Negative False ative ative positive

The results of Examples 2-1 to 2-2 and 3-1 to 3-2, and ComparativeExamples 2-1 to 2-2 and 3-1 to 3-3 indicate that compared with the casewhere the inside wall was not treated at all, the polypropylene bloodsampling tube and polyethylene terephthalate blood sampling tubecarrying only the blood coagulation accelerator silica spray-coated onthe inside wall (Comparative Example2-2 and Comparative Example 3-1,respectively) suffer from more remarkable clot deposits and fail topermit recovery of serum, the use of the blood componentdeposition-preventing agent of the invention in combination with silicainsures good serum separability without inhibiting thecoagulation-accelerating action of silica.

Furthermore, the results of Examples 3-1 to 3-2 and Comparative Examples3-1 to 3-3 indicate that whereas the blood componentdeposition-preventing agent of the present invention does not inducefalse positive tests for HBs-Ab and Free T4, the conventional nonionicsurfactant give false positive tests.

Example 4-1

Using a vinylpyrrolidone-vinyl acetate random copolymer (manufactured byBASF, Luviskol(tradename) VA28, vinyl acetate content ca 84 mol %) asthe blood component deposition-preventing agent, a methanolic solutionof 0.02 weight % concentration was prepared, spray-coated on the insidewall of a polyethylene terephthalate (PET) tube of 10 ml capacity (16 mmin. dia.×100 mm long) and air-dried. The coating amount per unit area ofthe inside wall was 5×10⁻⁷ g/cm² as vinylpyrrolidone-vinyl acetaterandom copolymer.

Then, a liquid dicyclopentadiene (DCPD) resin (manufactured by Exon(tradename) ECR-327), which is a plasma separator, was mixed with finelydivided silica (manufactured by Japan Aerosil Co., (tradename) AerosilA-200) under agitation to prepare a composition with a specific gravityof 1.05 and 1.2 g of the composition was put in the tube. Then, 120 U ofheparin sodium as the blood anticoagulant was further accommodated toprovide a blood test ware.

Fresh human blood, 8 ml, was put in the above blood test tube and thetube was stoppered and turned upside down 3 times for blending. The tubewas then allowed to sit at 20° C. for 10 minutes and, thereafter,centrifuged at 3000 rpm for 5 minutes to observe the separability ofplasma. At the same time, ½ of the supernatant plasma was pipetted foruse as a sample immediately after centrifugation.

Further, the blood test ware after centrifugation was stored at 40° C.for 24 hours and the supernatant plasma was pipetted to provide a24-hour storage sample.

Using the above sample after centrifugation and the 24-hour storagesample, lactic dehydrogenase (LDH), creatine kinase (CPK) and potassium(K) concentrations were determined. The results are shown in Table 1-6.The measured values presented in Table 1-6 are the relative values withthe value found for the sample immediately after centrifugation beingtaken as 100.

Comparative Example 4-1

A glass tube of 10 ml capacity (16 mm in. dia.×100 mm long) was chargedwith 120 U of heparin sodium as anticoagulant to provide a blood testware (neither the vinylpyrrolidone-vinyl acetate random copolymer northe plasma separator was used).

The performance evaluation of this blood test ware was carried out inthe following manner. Thus, whereas the performance evaluation inExample 4-1 was made by “storing the blood test ware aftercentrifugation at 4° C. and pipetting the supernatant plasma again after24 hours for use as a 24-hour storage sample”, “the whole amount of theplasma was recovered from the blood test ware after centrifugation and ½of the plasma was taken as a sample immediately after centrifugation,while the remainder was transferred to another glass tube and stored at40° C. for 24 hours for use as a 24-hour storage sample”. Otherwise theperformance evaluation was made in the same manner as described inExample 4-1. The results are shown in Table 1-6.

Comparative Example 4-2

Except that the vinylpyrrolidone-vinyl acetate random copolymer was notused, the procedure of Example 4-1 was otherwise repeated to provide ablood test ware (this ware contained the plasma separator and bloodanticoagulant). The performance evaluation of this blood test ware wascarried out in the same manner as described in Example 4-1. The resultsare shown in Table 1-6.

TABLE 1-6 Separability Measured value of serum LDH CPK K Example 4-1Good 100 100 105 Comparative Example 4-1 Good 100 100 100 ComparativeExample 4-2 Poor 120 130 110

Example 5-1

Using a vinylpyrrolidone-vinyl acetate random copolymer (manufactured byBASF, Luviskol (tradename) VA64, vinyl acetate content ca 46 mol %) asthe blood component deposition-preventing agent and heparin lithium asthe blood anticoagulant, a mixed solution containing 1.0 weights and4000 IU/ml, respectively, of these substances in pure water wasprepared. About 25 μl of this solution was spray-coated on the insidewall of a polyethylene terephthalate) blood sampling tube of 7 mlcapacity and dried in an air-current dryer at 60° C. Then, about 1 g ofa pasty serum/plasma separator (manufactured by Sekisui Kagaku KogyoKabusttiki Kaisha, S-Collect (tradename)) was introduced into the bottomportion of the dried blood sampling tube to provide a blood test ware,

This blood test ware was charged with 6 ml of fresh rabbit blood, turnedupside down for through blending, and centrifuged (25° C.) at 1300 G for5 minutes to visually evaluate the separability of plasma. Immediatelythen, about one-half volume of the plasma was transferred to a cleanhard glass test tube (a sample for initial baseline values), while theremainder as contained in the poly(ethylene terephthalate) tube wasstored in the refrigerator at 4° C. for 24 hours.

After 24 hours, the remaining plasma was transferred to a clean hardglass tube (a 24-hour storage sample for 24-hour values). Using theabove two plasma samples taken in hard glass test tubes, the influenceson blood chemistry parameters (LDH as an enzyme and K as an electrolyte)were investigated. The results are shown in Table 1-7.

Example 5-2

Except that a vinyl alcohol-vinyl acetate random copolymer (UnitikaLtd., Unitika Poval UMR-30L (tradename), vinyl acetate content ca. 60mol %) was used in lieu of said vinylpyrrolidone-vinyl acetate randomcopolymer, the procedure of Example 5-1 was otherwise repeated toprovide a blood test ware and using the ware, tests were performed as inExample 5-1. The results are shown in Table 1-7.

Comparative Example 5-1

Except that the use of vinylpyrrolidone-vinyl acetate random copolymeras the blood component deposition-preventing agent was omitted, theprocedure of Example 5-1 was otherwise repeated to provide a blood testware (the heparin lithium and serum/plasma separator were used). Usingthis blood test ware, the same tests were performed as in Example 5-1.The results are shown in Table 1-7.

Comparative Example 5-2

The inside wall of a clean hard glass blood sampling tube of 7 mlcapacity was spray-coated with about 25 μl of a solution containing 4000IU/ml of beparin lithium in pure water and dried in an air-current dryerat 60° C. to prepare a blood test ware (the serum/plasma separator wasnot introduced). This blood test tube was charged with 6 ml of freshrabbit blood, turned upside down a sufficient times for blending, andcentrifuged (25° C.) at 1300 G for 5 minutes to evaluate plasmaseparability after centrifugation as in Example 5-1. Immediately thenthe whole amount of plasma was transferred to another clean hard glasstest tube (a sample for initial baseline values) and stored in therefrigerator at 4° C. for 24 hours. After 24 hours, the same tests as inExample 5-1 were performed. The results are shown in Table 1-7.

TABLE 1-7 Example Comparative Example 5-1 5-2 5-1 5-2 Adhesion of bloodNone None Adhesion of None components after platelets centrifugationLDH(IU/L) Initial 123 129 120 123 After 24 hr 119 121 305 — of standingK Initial 4.0 4.0 4.1 4.0 (mEq/L) After 24 hr 4.3 4.5 5.3 — of standing

The results of Examples 5-1 to 5-2 and Comparative Examples 5-1 to 5-2indicate that the poly(ethylene terephthalate) blood sampling tubecarrying a spray-coated anticoagulant heparin salt on its inside wallshowed deposits of platelets and because of the leakage of LDH and Kfrom the platelets during storage at 40° C., the corresponding parametervalues are elevated. In Contrast, the blood componentdeposition-preventing agent of the present invention effectivelyprevented deposition of platelets, thus insuring stability of suchparameter values. Similar findings were also obtained between Example4-1 and Comparative Examples 4-1 and 4-2.

Example 6-1

Using a vinylpyrrolidone-vinyl acetate random copolymer (manufactured byBASF, Luviskol TM VA64, vinyl acetate content ca 46 mol %) as the bloodcomponent deposition-preventing agent and the same finely divided silicaas used in Example 2-1 as the blood coagulation accelerator, asuspension containing 1.0 weight % and 2.0 weight % of the respectivesubstances in methanol was prepared. This suspension was used to coatpolystyrene pellets having a diameter of about 3 mm in anexplosion-proof air-current dryer at 60° C. under agitation and dried toprovide a blood component deposition-preventing matrix. The amounts ofdeposition of the respective substances on the surface of pellets wereabout 2×10⁻⁵ g/cm² for vinylpyrrolidone-vinyl acetate random copolymerand about 7×10⁻⁷ g/cm² for finely divided silica.

A clean hard glass blood sampling tube of 10 ml capacity was chargedwith 0.6 g of the above blood component deposition-preventing matrix toprovide a blood test ware. This blood test ware was charged with 4 ml offresh rabbit blood and allowed to sit at the room temperature of 23 to25° C. After measurement of blood coagulation time, the sample wascentrifuged (25° C.) at 1300° C. for 5 minutes to visually evaluateserum separability and the serum yield was determined. As a result,although the pellets were found scatteredly buried near the head of theclot, a clean serum without showing signs of hemolysis was obtained. Thecoagulation time and serum yield values are shown in Table 1-8 .

Comparative Example 6-1

Except that the use of vinylpyrrolidone-vinyl acetate random copolymerwas omitted, the procedure of Example 6-1 was otherwise repeated toprepare silica powder-coated polystyrene pellets. The amount ofdeposition of finely divided silica was about 5×10⁻⁵ g/cm². A clean hardglass blood sampling tube of 10 ml capacity was filled with 0.6 g of thecoated polystyrene pellets to provide a blood test ware. This blood testware was charged with 4 ml of fresh rabbit blood and the bloodcoagulation time, serum separability, and serum yield were determined asin Example 6-1. As a result, the pellets were found buried near the headof the clot as in Example 6-1. The serum yield was as good as thatobtained in Example 6-1 but marked hemolysis was observed. Thecoagulation time and serum yield values are presented in Table 1-8.

Comparative Example 6-2

A clean hard glass blood sampling tube of 10 ml capacity was chargedwith 0.6 g of uncoated polystyrene pellets to provide a blood test ware.This blood test ware was filled with 4 ml of fresh rabbit blood and theblood coagulation time, serum separability, and serum yield weredetermined as in Example 6-1. As a result, the pellets were found buriednear the head of the clot as in Example 6-1. The serum yield was as goodas that obtained in Example 6-1 but marked hemolysis was observed. Thecoagulation time and serum yield values are presented in Table 1-8.

TABLE 1-8 Blood coagulation Separability Serum yield time (min.) ofserum (ml) Example 6-1 20 Good 2.2 Comparative 20 Very remarkable 2.0Example 6-1 hemolysis Comparative 45 Remarkable 2.1 Example 6-2hemolysis

Examples 7-1 to 7-12 and Comparative Example 7-1 Confirmation ofAntimicrobial Activity

Blood coagulation accelerator suspensions in pure water were preparedaccording to the formulas shown in Table 2-1l In Table 2-1, Bactekillermeans BM103A manufactured by Kanebo, Ltd., Ice means NAZ320 manufacturedby Catalysts & Chemicals Industries Co., Ltd., Rasap means AN600manufactured by Rasa Industries, Ltd., and Novalon means AG300manufactured by Toa Gosei Chemical Industry Co., Ltd.

Separately, a suspension of silica powder (1.0%) andpolyvinylpyrrolidone (2.0%) in pure water was prepared and exposed tointerior air for microbial contamination for use as an inoculum. Usingthe suspensions according to Table 2-1 as they were and thecorresponding suspensions obtained by inoculation with seedmicroorganisms and subsequent 10 hours of agitation, bacterial andfungal culture assays were carried out in the routine manner. The mediaand cultural conditions were as follows. For bacteria, culture wascarried out using Standard Method Agar at 30° C. for 3 days. Theinoculum size of each suspension was set at 0.3 ml. For fungi, culturewas carried out using Potato Dextrose Agar at 25° C. for 5 days. Theinoculum size of each suspension for fungal assays was 0.3 ml. Thefrequency of colonies on each medium was recorded. the results for themedia used in the culture of bacteria and fungi are shown in Table 2-2and Table 2-3, respectively. In each table, − means no colony formationand +++ means formation of many colonies.

Examples 7-13 to 7-20 Confirmation of Influences on Blood Coagulation

Blood coagulation accelerator suspensions in pure water were preparedaccording to the formulas shown in Table 2-4. About 50 μl of eachsuspension was spray-coated on a poly(methyl methacrylate) bloodsampling tube of 10 ml capacity and dried in an air-current dryer at 60°C. Then, fresh rabbit blood, 3 ml, was drawn into each blood samplingtube and allowed to stand at the room temperature of 23 to 25° C. Thetime till the blood lost fluidity and the serum began to seep out wasmeasured as blood coagulation time. The results are shown in Table 2-5.

Examples 7-11 to 7-28 Confirmation of Influences on Blood ExaminationParameter Values

After completion of the evaluation described in Examples 7-13 to 7-20,each sample was centrifuged at 1800 G for 5 minutes to recover theserum. Using the serum, influence on representative biochemicalparameters (GOT and ALP as enzymes, TG, PL and T-CHO as lipids, and Na,K, Cl, Mg, and Ca as electrolytes, and Fe and Cu as metals) wereinvestigated. The results are presented in Table 2-6. In Table 2-6, theblood sampling tubes of Example 7-21 to 7-28 correspond to the bloodsampling tubes of examples 7-13 to 7-20, respectively.

Comparative Examples 7-2 to 7-4

A blood coagulation accelerator suspension in pure water which containedno antimicrobial composition was prepared according to the formula shownin Table 2-4. About 50 μl of this suspension was spray-coated on apoly(methyl methacrylate) blood sampling tube of 10 ml capacity anddried in an air-current dryer at 60° C. (Comparative Example 7-2).Separately, a 10 ml hard glass blood sampling tube not spray-coated withthe blood coagulation accelerator (Comparative Example 7-3) and asimilar poly(methyl methacrylate) blood sampling tube (ComparativeExample 7-4) were provided. Fresh rabbit blood was drawn into these 3different blood sampling tubes and the blood coagulation times weredetermined in the manner described in Example 7-13. The results areshown alongside the results for Examples 7-13 to 7-20 in Table 2-5.

Comparative Example 7-5 to 7-7

After completion of the evaluation described in Comparative Examples 7-2to 7-4, each sample was centrifuged at 1800 G for 5 minutes to recoverthe serum and, using the serum, the influences on biochemical parametervalues were investigated. The results are shown alongside the resultsfor Examples 7-21 to 7-28 in Table 2-6. In Table 2-6, the blood samplingtubes of Comparative Examples 7-5 to 7-7 correspond to the bloodsampling tubes of Comparative Examples 7-2 to 7-4, respectively.

It will be apparent from the results shown in Table 2-2 and Table 2-3that when a germ-free coagulation accelerator suspension immediatelyafter preparation was contaminated by microorganisms, the microorganismswere ready to multiply in the absence of an antimicrobial composition(Comparative Example 7-1) but in cases where an antimicrobialcomposition was contained (Examples 7-1 to 7-12), invariably no colonieswere detected, indicating that the invading microorganisms weredestroyed.

The results shown in Table 2-5 indicate that it took more than 1 hourfor the blood to be coagulated in the plain poly(methyl methacrylate)blood sampling tube not treated with a coagulation accelerator(Comparative Example 7-4) but coagulation was completed within 20 to 30minutes in the presence of the blood coagulation accelerator accordingto the present invention (Examples 7-13 to 7-20). The latter result isfully comparable not only to the result with the conventionalcoagulation accelerator in the absence of an antimicrobial composition(Comparative Example 7-2) but also to the result with the hard glassblood sampling tube (Comparative Example 7-3), indicating that theantimicrobial composition of the present invention does not interferewith blood coagulation but rather has positive coagulation acceleratingactivity.

The results shown in Table 2-6 indicate that none of the antimicrobialcompositions had any remarkable confounding effects on blood parametervalues.

TABLE 2-1 Antimicrobial Coagulation Binder (%) composition (%)accelerator (%) Polyvinyl- Bactekiller Ice Rasap Novarone Silicapyrrolidone Example 7-1 0.10 — — — 1.0 2.0 Example 7-2 0.50 — — — 1.02.0 Example 7-3 1.00 — — — 1.0 2.0 Example 7-4 — 0.10 — — 1.0 2.0Example 7-5 — 0.50 — — 1.0 2.0 Example 7-6 — 1.00 — — 1.0 2.0 Example7-7 — — 0.10 — 1.0 2.0 Example 7-8 — — 0.50 — 1.0 2.0 Example 7-9 — —1.00 — 1.0 2.0 Example 7-10 — — — 0.10 1.0 2.0 Example 7-11 — — — 0.501.0 2.0 Example 7-12 — — — 1.00 1.0 2.0 Comparative — — — — 1.0 2.0Example 7-1

TABLE 2-2 Example Comparative Example 7-1 7-2 7-3 7-4 7-5 7-6 7-7 7-87-9 7-10 7-11 7-12 7-1 Suspension − − − − − − − − − − − − − Inoculatedsuspension − − − − − − − − − − − − +++

TABLE 2-3 Example Comparative Example 7-1 7-2 7-3 7-4 7-5 7-6 7-7 7-87-9 7-10 7-11 7-12 7-1 Suspension − − − − − − − − − − − − − Inoculatedsuspension − − − − − − − − − − − − +++

TABLE 2-4 Antimicrobial Coagulation Binder (%) composition (%)accelerator (%) Polyvinyl- Bactekiller Ice Rasap Novarone Silicapyrrolidone Example 7-13 0.10 — — — 0.9 2.0 Example 7-14 1.00 — — — —2.0 Example 7-15 — 0.10 — — 0.9 2.0 Example 7-16 — 1.00 — — — 2.0Example 7-17 — — 0.10 — 0.9 2.0 Example 7-18 — — 1.00 — — 2.0 Example7-19 — — — 0.10 0.9 2.0 Example 7-20 — — — 1.00 — 2.0 Comparative — — —— 1.0 2.0 Example 7-2 to 7-4

TABLE 2-5 Comparative Example Example 7-13 7-14 7-15 7-16 7-17 7-18 7-197-20 7-2 7-3 7-4 Blood coagulation 30 30 20 20 25 30 20 30 20 25 85 time(min.)

TABLE 2-6 Comparative Example Example 7-21 7-22 7-23 7-24 7-25 7-26 7-277-28 7-5 7-6 7-7 GOT (IU/l) 167 164 168 165 159 171 174 162 169 173 165ALP (IU/l) 101 102 100 102 98 100 101 100 99 101 98 TG (mg/dl) 56 53 5652 53 55 54 50 53 54 52 PL (mg/dl) 51 50 50 49 49 50 49 49 50 50 49T-CHO (mg/dl) 16 16 16 16 16 16 16 16 16 16 16 Na (mEq/l) 146 149 148146 148 148 146 150 149 146 147 K (mEq/l) 5.0 5.0 5.2 5.0 5.0 5.3 4.95.3 5.1 4.9 4.9 Cl (mEq/l) 98 101 100 101 100 102 98 101 98 100 100 Mg(mg/dl) 4.1 4.1 4.0 4.2 4.0 4.1 4.1 4.0 4.1 4.0 4.0 Ca (mg/dl) 13.9 13.713.9 13.5 13.9 14.1 14.2 14.1 13.9 14.2 14.1 Fe (μg/dl) 217 213 218 216215 211 213 214 208 213 215 Cu (μg/dl) 152 145 146 146 150 147 145 147150 145 147

Example 1

Using a polyvinylpyrrolidone (manufactured by BASF, tradename LuviskolK80, K value 80, weight average molecular weight ca 800000), adispersion of 0.1 weight % concentration in methanol was prepared. Thisdispersion was spray-coated on the inside wall of a poly(ethyleneterephthalate) (PET) tube of 10 ml capacity (16 mm in. dia.×100 mm long)and air-dried. The coating amount per unit area of the inside wall ofthe tube was 2×10⁻⁶ g/cm² as polyvinylpyrrolidone.

Further, as the plasma separator, dicyclopentadiene (DCPD) resin(manufactured by Exon tradename ECR-327) was mixed with finely dividedsilica (manufactured by Japan Aerosil Co., tradename Aerosil A-200) toprepare a composition with a specific gravity of 1.05 and 1.2 g of thecomposition was introduced into the above tube. Then., 120 U of heparinsodium as blood anticoagulant was further introduced to provide a bloodtest ware.

Example 2

Using a polyvinylpyrrolidone (manufactured by BASF, tradename LuviskolK90, K value 90, weight average molecular weight ca 1100000), adispersion of 0.02 weight % concentration in methanol was prepared andthis dispersion was spray-coated on the inside wall of a PET tubesimilar to the tube used in Example 1 and air-dried. The coating amountper unit area of the inside wall was 6×10⁻⁷ g/cm² aspolyvinylpyrrolidone. Thereafter, the plasma separator and the bloodanticoagulant were introduced as in Example 1 to provide a blood testware.

Comparative Example 1

A glass tube of 10 ml capacity (16 mm in. dia.×100 mm long) was chargedwith 120 U of heparin sodium as a blood anticoagulant to provide a bloodtest ware (neither polyvinylpyrrolidone nor a plasma separator wasused).

Comparative Example 2

Except that the use of polyvinylpyrrolidone was omitted, the procedureof Example 1 was repeated to provide a blood test ware (the plasmaseparator and the anticoagulant were. used).

Performance Evaluation

The relative performance of the blood test wares according to Examples 1and 2-and Comparative Example 2 was evaluated as follows.

Fresh human blood, 8 ml, was drawn into each blood test ware andstoppered tight. The ware was turned upside down 3 times for blending,then allowed to sit at 20° C. for 10 minutes, and centrifuged at 3000rpm for 5 minutes to assess plasma separability. At the same time, ½ ofthe supernatant plasma was quickly pipetted as a sample immediatelyafter centrifugation.

The same centrifuged blood test ware was stored at 4° C. for 24 hoursand the supernatant plasma was pipetted again for use as a 24-hourstorage sample.

Using the above sample immediately after centrifugation and 24-hourstorage sample, lactic dehydrogenase (LDH), creatine kinase (CPK) andpotassium (K) were assayed immediately after centrifugation for theformer sample and at 24 hours after centrifugation for the latter24-hour storage sample. The values found are presented in Table 1. Theassay method was the lactic substrate method for LDH, the creatinephosphate substrate method for CPK, and flame photometry for K. Themeasured values shown in Table 1 are relative values with the valuesfound for the sample immediately after centrifugation being taken as100.

The performance evaluation of the blood test ware of Comparative Example1 was carried out in the same manner as above, except that whereas theperformance evaluation in Example 4-1 was made by “storing the bloodtest ware after centrifugation at 4° C. and pipetting the supernatantplasma again after 24 hours for use as a 24-hour storage sample”, “theplasma recovered from the blood test ware immediately aftercentrifugation was transferred to another glass tube and stored at 4° C.for 24 hours and the supernatant plasma was used as a 24-hour storagesample”.

TABLE 1 Separability Measured value of serum LDH CPK K Example 1 Good105 105 100 Example 2 Good 105 105 105 Comparative Example 1 Good 100100 100 Comparative Example 2 Poor 120 130 110

Examples 3 to 8 and Comparative Examples 3 and 4

Aqueous solutions each containing both polyvinylpyrrolidone and heparinsodium at the concentrations indicated in Table 2 were prepared. Each ofthese aqueous solutions was spray-coated on the inside wall of a PETtube similar to the one used in Example 1 and dried. The coating amountsof polyvinylpyrrolidone and heparin sodium per unit area of the insidewall are shown in Table 2.

The vinylpyrrolidone polymers shown in table 2 are invariably theproducts of BASF and the correspondence between the indicated weightaverage molecular weight and the tradename are: ca 50000=Luviskol K30,ca 350000 =Luviskol K60, ca 800000=Luviskol K80, and ca 1100000=LuviskolK90.

Then, the plasma separator was introduced as in Example 1 to provideblood test wares.

Comparative Example 5

An aqueous solution containing 4500 U/ml of heparin sodium, 20 μl , as ablood anticoagulant was spray-coated on the inside wall of a glass tubeof 10 ml capacity (16 mm in. dia.×100 mm long) and dried to provide ablood test ware. The coating amount, as heparin sodium, per unit area ofthe inside wall of the tube is shown in Table 2.

Performance Evaluation

The relative performance of the blood test wares according to Examples 3to 8 and Comparative Examples 3 to 5 was evaluated as follows.

Fresh human blood, 3 ml, was drawn into each blood test ware andstoppered tight. The ware was turned upside down 3 times for blending,then allowed to sit at 20° C. for 10 minutes, and centrifuged at 3000rpm for 5 minutes to assess plasma separability. The results are shownin Table 3.

Then, ½ of the supernatant plasma was taken, transferred to a freshclean hard glass blood sampling tube, and stored frozen as a sampleimmediately after centrifugation.

The remaining plasma in the blood test ware was stored as it was at 4°C. for 24 hours. Then, it was transferred to a fresh clean hard glassblood sampling tube and stored frozen for use as a 24-hour storagesample.

As to the blood test ware prepared in Comparative Example 5, the plasmaseparability was evaluated and, then, the whole amount of supernatantplasma was transferred to a fresh clean hard glass blood sampling tubeand stored frozen for use as a sample immediately after centrifugation.

All the frozen samples were thawed 48 hours later and the concentrationsof lactic dehydrogenase (LDH), creatine kinase (CPK) and potassium (K)were determined as in Example 1. The results are presented in Table 3.The values given in Table 3 are the measured values.

TABLE 2 Polyvinylpyrrolidone Heparin sodium Amount of Amount of Weightaverage Concentration deposits Concentration deposits K molecular weight(wt %) (g/cm²) (U/ml) (U/cm²) Example 3 60 ca 35 × 10⁴ 0.1 3 × 10⁻⁷ 45001.4 Example 4 60 ca 35 × 10⁴ 5.0 2 × 10⁻⁵ 4500 1.9 Example 5 80 ca 80 ×10⁴ 0.1 1 × 10⁻⁶ 4500 4.8 Example 6 80 ca 80 × 10⁴ 5.0 4 × 10⁻⁵ 4500 3.4Example 7 90 ca 110 × 10⁴  0.1 9 × 10⁻⁷ 4500 3.8 Example 8 90 ca 110 ×10⁴  5.0 7 × 10⁻⁵ 4500 6.3 Comparative Example 3 30  ca 5 × 10⁴ 0.1 4 ×10⁻⁷ 4500 1.9 Comparative Example 4 30  ca 5 × 10⁴ 5.0 4 × 10⁻⁵ 4500 3.4Comparative Example 5 — — — — 4500 2.0

TABLE 3 Separa- LDH (IU/l) CPK (IU/l) K (mEq/l) bility ImmediatelyImmediately Immediately of after after after plasma separation After 24hr separation After 24 hr separation After 24 hr Example 3 ∘ 200 200 103 93 3.8 3.9 Example 4 ∘ 205 201 101 102 3.8 4.1 Example 5 ∘ 198 204 105104 3.9 4.0 Example 6 ∘ 205 210 102 105 3.9 4.0 Example 7 ∘ 208 201 109107 3.9 4.0 Example 8 ∘ 200 201  99 102 3.8 4.0 Comparative x 204 238 98 119 4.0 4.9 Example 3 Comparative x 201 254 100 123 3.9 5.2 Example4 Comparative ∘ 204 — 101 — 4.0 — Example 5

In the plasma separability column of Table 3, ∘ denotes good and Xdenotes marked deposition of platelets. Examples 9 to 24 and ComparativeExamples 6 to 9

The formulas for the compositions coated on the inside walls of bloodsampling tubes are shown in Table 4. In Table 4, Luviskol K80 means agrade of polyvinylpyrrolidone (κ value 80, weight average molecularweight ca 800000) manufactured by BASF, Luviskol K90 means a grade ofpolyvinylpyrrolidone (K value 90, weight average molecular weight ca1100000) manufactured by BASF, EDTA2K means dipotassiumethylenediaminetetraacetate (manufactured by Wako Pure ChemicalIndustries, Ltd., reagent grade), Heparin Li means a reagent gradeproduct of Sigma Chemical Company, and PMMA powder means a poly(methylmethacrylate) powder having a particle diameter of about 50 μm (MB-50,manufactured by Sekisui Kagaku Kogyo Kabushaiki Kaisha), and Cellulosepowder means a finely divided cellulose having a particle diameter ofabout 20 μm (reagent grade, manufactured by Aldrich Chemical Company,Inc.).

TABLE 4 Polyvinyl- Blood Finely divided pyrrolidone anticoagulantinsoluble powder Example 9 Luviskol K80 (0.1 w/w %) EDTA2K (10 w/w %)PMMA powder (0.1 w/w %) Example 10 Luviskol K80 (0.1 w/w %) EDTA2K (10w/w %) Cellulose powder (0.1 w/w %) Example 11 Luviskol K80 (0.1 w/w %)EDTA2K (10 w/w %) PMMA powder (5.0 w/w %) Example 12 Luviskol K80 (0.1w/w %) EDTA2K (10 w/w %) Cellulose powder (5.0 w/w %) Example 13Luviskol K90 (0.1 w/w %) EDTA2K (10 w/w %) PMMA powder (5.0 w/w %)Example 14 Luviskol K90 (0.1 w/w %) EDTA2K (10 w/w %) Cellulose powder(5.0 w/w %) Example 15 Luviskol K90 (0.1 w/w %) EDTA2K (10 w/w %) PMMApowder (0.1 w/w %) Example 16 Luviskol K90 (0.1 w/w %) EDTA2K (10 w/w %)Cellulose powder (0.1 w/w %) Example 17 Luviskol K80 (0.1 w/w %) HeparinLi (1000 IU/ml) PMMA powder (0.1 w/w %) Example 18 Luviskol K80 (0.1 w/w%) Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w %) Example 19Luviskol K80 (0.1 w/w %) Heparin Li (1000 IU/ml) PMMA powder (5.0 w/w %)Example 20 Luviskol K80 (0.1 w/w %) Heparin Li (1000 IU/ml) Cellulosepowder (5.0 w/w %) Example 21 Luviskol K90 (0.1 w/w %) Heparin Li (1000IU/ml) PMMA powder (0.1 w/w %) Example 22 Luviskol K90 (0.1 w/w %)Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w %) Example 23 LuviskolK90 (0.1 w/w %) Heparin Li (1000 IU/ml) PMMA powder (5.0 w/w %) Example24 Luviskol K90 (0.1 w/w %) Heparin Li (1000 IU/ml) Cellulose powder(5.0 w/w %) Comparative Example 6 Luviskol K80 (0.1 w/w %) EDTA2K (10w/w %) Not used Comparative Example 7 Luviskol K90 (0.1 w/w %) EDTA2K(10 w/w %) Not used Comparative Example 8 Luviskol K80 (0.1 w/w %)Heparin Li (1000 IU/ml) Not used Comparative Example 9 Luviskol K90 (0.1w/w %) Heparin Li (1000 IU/ml) Not used

Aqueous suspensions of the respective compositions according to Examples9 to 24 or aqueous solutions of the respective compositions according toComparative Examples 6 to 9 were prepared and about 30 μl of eachsuspension or solution was spray-coated on the inside wall of apoly(ethylene terephthalate) blood sampling tube of 7 ml capacity. Thetubes were then allowed to sit in the upright position at 600° C.overnight to dry. Then, the retentivity of each composition on theinside wall of the tube was visually evaluated. The results are shown inTable 5. The results of these Examples and Comparative Examples indicatethat the concomitant use of a finely divided insoluble powder results ina marked improvement in the retentivity of the suspension or solution onthe inside wall of the tube.

TABLE 5 Example 9 10 11 12 13 14 15 16 Retentivity Good Good Good GoodGood Good Good Good Example 17 18 19 20 21 22 23 24 Retentivity GoodGood Good Good Good Good Good Good Comparative Example 6 7 8 9Retentivity Marked Marked Marked Marked sagging sagging sagging sagging

Then, a plasma separator (S-Collect, Sekisui Kagaku Kogyo KabushikiKaisha), 0.9 g/tube, was introduced into all the blood sampling tube.

Then, 3 ml/tube of fresh rabbit blood was drawn into all the bloodsampling tubes, which were then inverted once for gentle bending. Thetubes were then allowed to sit at the room temperature of about 23° C.for 30 minutes. Thereafter, each tube was centrifuged at 1300 G for 5minutes and immediately the deposition status of solid elements of bloodon the inside wall above the plasma separator partition was visuallyevaluated. The results are shown in Table 6. Because of the intentionalinsufficient blending, Comparative Examples in which most of thecomposition was buried in the plasma separator failed to show adequateanticoagulation effects with small amounts of the clot adhering to thetube in a ring fashion at the liquid level. In the Examples, however,satisfactory anticoagulation effects were obtained.

TABLE 6 Example 9 10 11 12 13 Retentivity No adhesion No adhesion Noadhesion No adhesion No adhesion Example 14 15 16 17 18 Retentivity Noadhesion No adhesion No adhesion No adhesion No adhesion Example 19 2021 22 23 24 Retentivity No adhesion No adhesion No adhesion No adhesionNo adhesion No adhesion Comparative Example 6 7 8 9 Retentivity A smallamount of A small amount of A small amount of A small amount of clotadhered in clot adhered in clot adhered in clot adhered in ring form atring form at ring form at ring form at liquid level liquid level liquidlevel liquid level

Then, from each of the blood sampling tubes, one-half of the plasma wastaken (initial baseline samples) and stored frozen at −20° C. in a 5 mlpoly(ethylene terephthalate) tube. The remaining one half of plasma wasstored as it was at 4° C. for 20 hours and, then, stored frozen in a 5ml poly(ethylene terephthalate) tube (20-hour storage samples) in thesame manner.

Forty-eight (48) hours after blood sampling, β-TG was assayed inExamples 9 to 16 and Comparative Examples 6 and 7 and LDH and K wereassayed in Examples 17 to 24 and Comparative Examples 8 and 9. Theresults are shown in Table 7. While β-TG is a substance contained at ahigh concentration in the platelets and LDH and K+ are substancescontained at high concentrations in both the platelets and erythrocytes,these substances are released when the blood cells are stimulated ordestroyed. In the Examples wherein a sufficient amount of thecomposition was retained on the inside wall of the tube, there was nodeposition of blood cells on the inside wall so that the measured valuesafter 24 hours of storage were not much different from the initialbaseline values prior to storage. In the Comparative Examples, in whichsmall amounts of the clot were found adhering to the inside wall, β-TGshowed higher values, both initially and after 20 hours of storage, thanin the Examples and LDU and K+showed upward changes in the course of 20hours' storage.

TABLE 7 β -TG (ng/ml) Sample after Initial 20 hr sample of standingExample 9 15 16 Example 10 15 16 Example 11 14 15 Example 12 13 15Example 13 17 15 Example 14 15 13 Example 15 17 17 Example 16 16 15Comparative Example 6 21 43 Comparative Example 7 25 47 LDH (IU/l) K(mEq/l) Sample after Sample after Initial 20 hr Initial 20 hr sample ofstanding sample of standing Example 17 105 107 4.1 4.1 Example 18 107105 4.1 4.1 Example 19 106 103 4.0 4.1 Example 20 103 105 4.0 4.1Example 21 105 107 4.0 4.1 Example 22 104 101 4.0 4.1 Exanple 23 103 1084.0 4.1 Example 24 107 105 4.0 4.1 Comparative Example 8 105 138 4.1 5.8Comparative Example 9 109 154 4.1 6.1

Examples 25 to 40 and Comparative Examples 10 to 13

The formulas for the compositions coated on the inside walls of bloodsampling tubes are shown in Table 8. In Table 8, VA64 means the randomcopolymer Luviskol VA64 manufactured by BASF (the monomer componentgiving a water-soluble homopolymer=46 mol % (40 weight %)), UMR-30Lmeans the random copolymer Unitika Poval UMR-30L manufactured by UnitikaLtd. (the monomer component giving a water-soluble homopolymer=60 ml %),EDTA2K means dipotassium ethylenediaminetetraacetate (Wako Pure ChemicalIndustries, Ltd., reagent grade), Heparin Li means the correspondingreagent grade manufactured by Sigma Chemical Company, PMMA powder meansa poly(methyl methacrylate) powder having a particle diameter of about50 μm (MB-50, manufactured by Sekisui Kagaku Kogyo Kabushiki Kaisha),and Cellulose powder means a finely divided cellulose having a particlediameter of about 20 μm (reagent grade, Aldrich Chemical Company, Inc.).

TABLE 8 Random Blood Finely divided copolymer anticoagulant insolublepowder Example 25 VA64 (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder (0.1w/w %) Example 26 VA64 (1.0 w/w %) EDTA2K (10 w/w %) Cellulose powder(0.1 w/w %) Example 27 VA64 (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder(5.0 w/w %) Example 28 VA64 (1.0 w/w %) EDTA2K (10 w/w %) Cellulosepowder (5.0 w/w %) Example 29 UMR-30L (1.0 w/w %) EDTA2K (10 w/w %) PMMApowder (5.0 w/w %) Example 30 UMR-30L (1.0 w/w %) EDTA2K (10 w/w %)Cellulose powder (5.0 w/w %) Example 31 UMR-30L (1.0 w/w %) EDTA2K (10w/w %) PMMA powder (0.1 w/w %) Example 32 UMR-30L (1.0 w/w %) EDTA2K (10w/w %) Cellulose powder (0.1 w/w %) Example 33 VA64 (1.0 w/w %) HeparinLi (1000 IU/ml) PMMA powder (0.1 w/w %) Example 34 VA64 (1.0 w/w %)Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w %) Example 35 VA64(1.0 w/w %) Heparin Li (1000 IU/ml) PMMA powder (5.0 w/w %) Example 36VA64 (1.0 w/w %) Heparin Li (1000 IU/ml) Cellulose powder (5.0 w/w %)Example 37 UMR-30L (1.0 w/w %) Heparin Li (1000 IU/ml) PMMA powder (0.1w/w %) Example 38 UMR-30L (1.0 w/w %) Heparin Li (1000 IU/ml) Cellulosepowder (0.1 w/w %) Example 39 UMR-30L (1.0 w/w %) Heparin Li (1000IU/ml) PMMA powder (5.0 w/w %) Example 40 UMR-30L (1.0 w/w %) Heparin Li(1000 IU/ml) Cellulose powder (5.0 w/w %) Comparative Example 10 VA64(1.0 w/w %) EDTA2K (10 w/w %) Not used Comparative Example 11 UMR-30L(1.0 w/w %) EDTA2K (10 w/w %) Not used Comparative Example 12 VA64 (1.0w/w %) Heparin Li (1000 IU/ml) Not used Comparative Example 13 UMR-30L(1.0 w/w %) Heparin Li (1000 IU/ml) Not used

Aqueous suspensions of the respective compositions according to Examples25 to 40 and aqueous solutions of the respective compositions accordingto Comparative Examples 10 to 13 were prepared and about 30 μl of eachsuspension or solution was spray-coated on the inside wall of apolyethylene terephthalate) blood sampling tube of 7 ml capacity. Thetubes were then allowed to sit in the upright position at 60° C.overnight to dry. Then, the retentivity of each composition on theinside wall of the tube was visually evaluated. The results are shown inTable 9. The results of these Examples and Comparative Examples indicatethat the concomitant use of a finely divided insoluble powder results ina marked improvement in the retentivity of the suspension or solution onthe inside wall of the tube.

TABLE 9 Example 25 26 27 28 29 30 31 32 Retentivity Good Good Good GoodGood Good Good Good Example 33 34 35 36 37 38 39 40 Retentivity GoodGood Good Good Good Good Good Good Comparative Example 10 11 12 13Retentivity Marked Marked Marked Marked sagging sagging sagging sagging

Then, a plasma separator (S-Collect, Sekisui Kagaku Kogyo KabushikiRaisha), 0.9 g/tube, was introduced into all the blood sampling tubes.

Then, 3 ml/tube of fresh rabbit blood was drawn into all the bloodsampling tubes, which were then inverted once for gentle blending. Thetubes were then allowed to sit at the room temperature of about 23° C.for 30 minutes. Thereafter, each tube was centrifuged at 1300 G for 5minutes and immediately the deposition status of solid elements of bloodon the inside wall above the plasma separator partition was visuallyevaluated. The results are shown in Table 10. Because of the intentionalinsufficient blending, the Comparative Examples in which most of thecomposition was buried in the plasma separator failed to show adequateanticoagulation effects with small amounts of the clot adhering to thetube wall in a ring fashion at the liquid level. In the Examples,however, satisfactory anticoagulation effects were obtained.

TABLE 10 Example 25 26 27 28 29 Retentivity No adhesion No adhesion Noadhesion No adhesion No adhesion Example 30 31 32 33 34 Retentivity Noadhesion No adhesion No adhesion No adhesion No adhesion Example 35 3637 38 39 40 Retentivity No adhesion No adhesion No adhesion No adhesionNo adhesion No adhesion Comparative Example 10 11 12 13 Retentivity Asmall amount of A small amount of A small amount of A small amount ofclot adhered in clot adhered in clot adhered in clot adhered in ringform at ring form at ring form at ring form at liquid level liquid levelliquid level liquid level

Then, from each blood sampling tube, one-half of the plasma was taken(initial baseline sample) and stored frozen at −20° C. in a 5 mlpoly(ethylene terephthalate) tube. The remaining one-half of plasma wasstored as it was at 4° C. for 20 hours and, then, stored frozen in a 5ml poly(ethylene terephthalate) tube (20-hour storage sample) in thesame manner.

Forty-eight (48) hours after blood sampling, β-TG was assayed inExamples 25 to 32 and Comparative Examples 10 and 11 and LDH and K wereassayed in Examples 33 to 40 and Comparative Examples 12 and 13. Theresults are shown in Table 11. While β-TG is a substance contained at ahigh concentration in the platelets and LDH and K+ are substancescontained at high concentrations in both the platelets and erythrocytes,these substances are released when the blood cells are stimulated ordestroyed. In the Examples wherein a sufficient amount of thecomposition was retained on the inside wall of the tube, there was nodeposition of blood cells on the inside wall so that the measured valuesafter 20 hours of storage were not much different from the initialbaseline values. In the Comparative Examples, in which small amounts ofthe clot were adhering to the inside wall, β-TG showed higher values,both initially and after 20 hours of storage, than in the Examples andLDH and K+ showed upward changes in the course of 20 hours' storage.

TABLE 11 β -TG (ng/ml) Sample after Initial 20 hr sample of standingExample 25 37 39 Example 26 33 31 Example 27 35 35 Example 28 35 37Example 29 38 35 Example 30 35 37 Example 31 34 33 Example 32 34 35Comparative Example 10 42 58 Comparative Example 11 39 62 LDH (IU/l) K(mEq/l) Sample after Sample after Initial 20 hr Initial 20 hr sample ofstanding sample of standing Example 33 121 123 3.9 3.9 Example 34 118120 3.9 3.9 Example 35 115 114 3.9 3.9 Example 36 117 117 3.8 3.9Example 37 117 116 3.9 3.9 Example 38 124 121 3.9 3.8 Example 39 119 1223.9 3.9 Example 40 121 124 3.9 3.8 Comparative Example 12 115 161 3.95.3 Comparative Example 13 117 159 3.9 5.1

Examples 41 to 56 and Comparative Examples 14 to 17

The formulas for the compositions coated on the inside walls of bloodsampling tubes are shown in Table 12. In Table 12, SH3749 means thepolyether-modified silicone oil nonionic surfactant SH3749 manufacturedby Toray-Dow Corning Silicone Co., Pluronic means thepolyoxyethylene-polyoxypropylene block copolymer nonionicsurfactant-Pluronic P75 manufactured by Asahi Denka, EDTA2K meansdipotassium ethylenediaminetetraacetate (Wako Pure Chemical Industries,Ltd., reagent grade), Heparin Li means the corresponding reagent grademanufactured by Sigma Chemical Company, PMMA powder means a poly(methylmethacrylate) powder having a particle diameter of about 50 μm (MB-50,manufactured by Sekisui Kagaku Kogyo Kabushiki Kaisha), and Cellulosepowder means a finely divided cellulose having a particle diameter ofabout 20 μm (reagent grade, Aldrich Chemical Company, Inc.).

TABLE 12 Nonionic Blood Finely divided surfactant anticoagulantinsoluble powder Example 41 SH3749 (0.3 w/w %) EDTA2K (10 w/w %) PMMApowder (0.1 w/w %) Example 42 SH3749 (0.3 w/w %) EDTA2K (10 w/w %)Cellulose powder (0.1 w/w %) Example 43 SH3749 (0.3 w/w %) EDTA2K (10w/w %) PMMA powder (5.0 w/w %) Example 44 SH3749 (0.3 w/w %) EDTA2K (10w/w %) Cellulose powder (5.0 w/w %) Example 45 Pluronic (0.3 w/w %)EDTA2K (10 w/w %) PMMA powder (5.0 w/w %) Example 46 Pluronic (0.3 w/w%) EDTA2K (10 w/w %) Cellulose powder (5.0 w/w %) Example 47 Pluronic(0.3 w/w %) EDTA2K (10 w/w %) PMMA powder (0.1 w/w %) Example 48Pluronic (0.3 w/w %) EDTA2K (10 w/w %) Cellulose powder (0.1 w/w %)Example 49 SH3749 (0.3 w/w %) Heparin Li (1000 IU/ml) PMMA powder (0.1w/w %) Example 50 SH3749 (0.3 w/w %) Heparin Li (1000 IU/ml) Cellulosepowder (0.1 w/w %) Example 51 SH3749 (0.3 w/w %) Heparin Li (1000 IU/ml)PMMA powder (5.0 w/w %) Example 52 SH3749 (0.3 w/w %) Heparin Li (1000IU/ml) Cellulose powder (5.0 w/w %) Example 53 Pluronic (0.3 w/w %)Heparin Li (1000 IU/ml) PMMA powder (0.1 w/w %) Example 54 Pluronic (0.3w/w %) Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w %) Example 55Pluronic (0.3 w/w %) Heparin Li (1000 IU/ml) PMMA powder (5.0 w/w %)Example 56 PIuronic (0.3 w/w %) Heparin Li (1000 IU/ml) Cellulose powder(5.0 w/w %) Comparative Example 14 SB3749 (0.3 w/w %) EDTA2K (10 w/w %)Not used Comparative Example 15 Pluronic (0.3 w/w %) EDTA2K (10 w/w %)Not used Comparative Example 16 SH3749 (0.3 w/w %) Heparin Li (1000IU/ml) Not used Comparative Example 17 Pluronic (0.3 w/w %) Heparin Li(1000 IU/ml) Not used

Aqueous suspensions of the respective compositions according to Examples41 to 56 and aqueous solutions of the respective compositions accordingto Comparative Examples 14 to 17 were prepared and about 30 μl of eachsuspension or solution was spray-coated on the inside wall of apoly(ethylene terephthalate) blood sampling tube of 7 ml capacity. Thetubes were then allowed to sit in the upright position at 60° C.overnight to dry. Then, the retentivity of each composition on theinside wall of the tube was visually evaluated. The results are shown inTable 13. The results of these Examples and Comparative Examplesindicate that the concomitant use of a finely divided insoluble powderresults in a marked improvement in the retentivity of the suspension orsolution on the inside wall of the tube.

TABLE 13 Example 41 42 43 44 45 46 47 48 Retentivity Good Good Good GoodGood Good Good Good Example 49 50 51 52 53 54 55 56 Retentivity GoodGood Good Good Good Good Good Good Comparative Example 14 15 16 17Retentivity Marked Marked Marked Marked sagging sagging sagging sagging

Then, a plasma separator (S-Collect, Sekisui Kagaku Kogyo KabushikiKaisha), 0.9 g/tube, was introduced into all the blood sampling tubes.

Then, 3 ml/tube of fresh rabbit blood was drawn into all the bloodsampling tubes, which were then inverted once for gentle blending. Thetubes were then allowed to sit at the room temperature of about 23° C.for 30 minutes. Thereafter, each tube was centrifuged at 1300 G for 5minutes and immediately the deposition status of cellular elements ofblood on the inside wall above the plasma separator partition wasvisually evaluated. The results are shown in Table 14. Because of theintentional insufficient blending, the Comparative Examples in whichmost of the composition was buried in the plasma separator failed toshow adequate anticoagulation effects with small amounts of the clotadhering to the tube in a ring fashion at the liquid level. In theExamples, however, satisfactory anticoagulation effects were obtained.

TABLE 14 Example 41 42 43 44 45 Retentivity No adhesion No adhesion Noadhesion No adhesion No adhesion Example 46 47 48 49 50 Retentivity Noadhesion No adhesion No adhesion No adhesion No adhesion Example 51 5253 54 55 56 Retentivity No adhesion No adhesion No adhesion No adhesionNo adhesion No adhesion Comparative Example 14 15 16 17 Retentivity Asmall amount of A small amount of A small amount of A small amount ofclot adhered in clot adhered in clot adhered in clot adhered in ringform at ring form at ring form at ring form at liquid level liquid levelliquid level liquid level

Then, from all the blood sampling tubes, one-half of the plasma wastaken (initial baseline samples) and stored frozen at −20° C. in a 5 mlpoly(ethylene terephthalate) tube. The remaining one-half of plasma wasstored as it was at 4° C. for 20 hours and, then, stored frozen in a 5ml poly(ethylene terephthalate) tube (20-hour storage samples) in thesame manner.

Forty-eight (48) hours after blood sampling, β-TG was assayed inExamples 41 to 48 and comparative Examples 14 and 15 and LDH and K wereassayed in Examples 49 to 56 and comparative Examples 16 and 17. Theresults are shown in Table 15. While β-TG is a substance contained at ahigh concentration in the platelets and LDH and K+ are substancescontained at high concentrations in both the platelets and erythrocytes,these substances are released when the blood cells are stimulated ordestroyed. In the Examples wherein a sufficient amount of thecomposition was retained on the inside wall of the tube, there was nodeposition of cellular blood components on the inside wall so that themeasured values after 20 hours of storage were not much different fromthe initial baseline values. In the Comparative Examples, in which smallamounts of the clot were adhering to the inside wall, β-TG showed highervalues, both initially and after 20 hours' storage, than in the Examplesand LDH and K+ showed upward changes in the course of 20 hours' storage.

TABLE 15 β -TG (ng/ml) Sample after Initial 20 hr sample of standingExample 41 41 43 Example 42 43 42 Example 43 45 44 Example 44 41 44Example 45 41 44 Example 46 45 41 Example 47 43 44 Example 48 44 47Comparative Example 14 58 75 Comparative Example 15 54 69 LDH (IU/l) K(mEq/l) Sample after Sample after Initial 20 hr Initial 20 hr sample ofstanding sample of standing Example 49 176 175 3.8 3.9 Example 50 180179 3.8 3.9 Example 51 181 175 3.8 3.9 Example 52 181 180 3.9 3.9Example 53 180 175 3.9 3.9 Example 54 178 175 3.9 3.9 Example 55 180 1783.9 3.9 Example 56 179 183 3.9 3.9 Comparative Example 16 183 256 4.05.9 Comparative Example 17 180 248 4.0 5.8

Examples 57 to 72 and Comparative Examples 18 to 25

The formulas for the compositions coated on the inside walls of bloodsampling tubes are shown in Tables 16 and 17. In Tables 16 and 17,Modified silicone oil means the carbinol-modified silicone oil SF8427manufactured by Toray-Dow Corning Silicone Co., Sorbitan monooleatemeans the corresponding reagent grade manufactured by Wako Pure ChemicalIndustries, Ltd., PVP means the polyvinylpyrrolidone K-30 manufacturedby BASF, PEG means the poly(ethylene glycol) with a number averagemolecular weight of about 10000 (manufactured by Aldrich ChemicalCompany, Inc., reagent grade), EDTA2K means dipotassiumethylenediaminetetraacetate (Wako Pure Chemical Industries, Ltd.,reagent grade), Heparin Li means the corresponding reagent grademanufactured by Sigma Chemical Company, PMMA powder means a poly(methylmethacrylate)powder having a particle diameter of about 50 μm (MB-50,manufactured by Sekisui Kagaku Kogyo Kabushiki Kaisha), and Cellulosepowder means a finely divided cellulose having a particle diameter ofabout 20 μm (reagent grade, Aldrich Chemical Company, Inc.).

TABLE 16 Practically insoluble Water-soluble Blood Finely dividedsubstance substance anticoagulant insoluble powder Example 57 Modifiedsilicone oil (1.0 w/w %) PVP (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder(0.1 w/w %) Example 58 Modified silicone oil (1.0 w/w %) PVP (1.0 w/w %)EDTA2K (10 w/w %) Cellulose powder (0.1 w/w %) Example 59 Modifiedsilicone oil (1.0 w/w %) PEG (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder(5.0 w/w %) Example 60 Modified silicone oil (1.0 w/w %) PEG (1.0 w/w %)EDTA2K (10 w/w %) Cellulose powder (5.0 w/w %) Example 61 Sorbitanmonooleate (1.0 w/w %) PVP (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder(5.0 w/w %) Example 62 Sorbitan monooleate (1.0 w/w %) PVP (1.0 w/w %)EDTA2K (10 w/w %) Cellulose powder (5.0 w/w %) Example 63 Sorbitanmonooleate (1.0 w/w %) PEG (1.0 w/w %) EDTA2K (10 w/w %) PMMA powder(0.1 w/w %) Example 64 Sorbitan monooleate (1.0 w/w %) PEG (1.0 w/w %)EDTA2K (10 w/w %) Cellulose powder (0.1 w/w %) Example 65 Modifiedsilicone oil (1.0 w/w %) PVP (1.0 w/w %) Heparin Li (1000 IU/ml) PMMApowder (0.1 w/w %) Example 66 Modified silicone oil (1.0 w/w %) PVP (1.0w/w %) Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w %) Example 67Modified silicone oil (1.0 w/w %) PEG (1.0 w/w %) Heparin Li (1000IU/ml) PMMA powder (5.0 w/w %) Example 68 Modified silicone oil (1.0 w/w%) PEG (1.0 w/w %) Heparin Li (1000 IU/ml) Cellulose powder (5.0 w/w %)Example 69 Sorbitan monooleate (1.0 w/w %) PVP (1.0 w/w %) Heparin Li(1000 IU/ml) PMMA powder (0.1 w/w %) Example 70 Sorbitan monooleate (1.0w/w %) PVP (1.0 w/w %) Heparin Li (1000 IU/ml) Cellulose powder (0.1 w/w%) Example 71 Sorbitan monooleate (1.0 w/w %) PEG (1.0 w/w %) Heparin Li(1000 IU/ml) PMMA powder (5.0 w/w %) Example 72 Sorbitan monooleate (1.0w/w %) PEG (1.0 w/w %) Heparin Li (1000 IU/ml) Cellulose powder (5.0 w/w%)

TABLE 17 Practically insoluble Water-soluble Blood Finely dividedsubstance substance anticoagulant insoluble powder Comparative Example18 Modified silicone oil (1.0 w/w %) PVP (1.0 w/w %) EDTA2K (10 w/w %)Not used Comparative Example 19 Modified silicone oil (1.0 w/w %) PEG(1.0 w/w %) EDTA2K (10 w/w %) Not used Comparative Example 20 Sorbitanmonooleate (1.0 w/w %) PVP (1.0 w/w %) EDTA2K (10 w/w %) Not usedComparative Example 21 Sorbitan monooleate (1.0 w/w %) PEG (1.0 w/w %)EDTA2K (10 w/w %) Not used Comparative Example 22 Modified silicone oil(1.0 w/w %) PVP (1.0 w/w %) Heparin Li (1000 IU/ml) Not used ComparativeExample 23 Modified silicone oil (1.0 w/w %) PEG (1.0 w/w %) Heparin Li(1000 IU/ml) Not used Comparative Example 24 Sorbitan monooleate (1.0w/w %) PVP (1.0 w/w %) Heparin Li (1000 IU/ml) Not used ComparativeExample 25 Sorbitan monooleate (1.0 w/w %) PEG (1.0 w/w %) Heparin Li(1000 IU/ml) Not used

Aqueous suspensions of the respective compositions according to Examples57 to 72 and aqueous solutions of the respective compositions accordingto Comparative Examples 14 to 17 were prepared and about 30 μl of eachsuspension or solution was spray-coated on the inside wall of apoly(ethylene terephthalate) blood sampling tube of 7 ml capacity. Thetubes were then allowed to sit in the upright position at 60° C.overnight to dry. Then, the retentivity of each composition on theinside wall of the tube was visually evaluated. The results are shown inTable 17. The results of these Examples and Comparative Examplesindicate that the concomitant use of a finely divided insolublesubstance results in a marked improvement in the retentivity of thesuspension or solution on the inside wall of the tube.

TABLE 18 Example 57 58 59 60 61 62 63 64 Retentivity Good Good Good GoodGood Good Good Good Example 65 66 67 68 69 70 71 72 Retentivity GoodGood Good Good Good Good Good Good Comparative Example 18 19 20 21Retentivity Marked Marked Marked Marked sagging sagging sagging saggingComparative Example 22 23 24 25 Retentivity Marked Marked Marked Markedsagging sagging sagging sagging

Then, a plasma separator (S-Collect, Sekisui Kagaku Kogyo KabushikiKaisha), 0.9 g/tube, was introduced into all the blood sampling tube.

Then, 3 ml/tube of fresh rabbit blood was drawn into all the bloodsampling tubes, which were then inverted once for gentle blending. Thetubes were then allowed to sit at the room temperature of about 23° C.for 30 minutes. Thereafter, each tube was centrifuged at 1300 G for 5minutes and immediately the deposition status of cellular elements ofblood on the inside wall above the plasma separator partition wasvisually evaluated. The results are shown in Table 18. Because ofintentional insufficient blending, the Comparative Examples in whichmost of the composition was buried in the plasma separator failed toshow adequate anticoagulation effects with small amounts of the clotadhering to the tube in a ring fashion at the liquid level. In theExamples, however, satisfactory anticoagulation effects were obtained.

TABLE 19 Example 57 58 59 60 61 Retentivity No adhesion No adhesion Noadhesion No adhesion No adhesion Example 62 63 64 65 66 Retentivity Noadhesion No adhesion No adhesion No adhesion No adhesion Example 67 6869 70 71 72 Retentivity No adhesion No adhesion No adhesion No adhesionNo adhesion No adhesion Comparative Example 18 19 20 21 Retentivity Asmall amount of A small amount of A small amount of A small amount ofclot adhered in clot adhered in clot adhered in clot adhered in ringform at ring form at ring form at ring form at liquid level liquid levelliquid level liquid level Comparative Example 22 23 24 25 Retentivity Asmall amount of A small amount of A small amount of A small amount ofclot adhered in clot adhered in clot adhered in clot adhered in ringform at ring form at ring form at ring form at liquid level liquid levelliquid level liquid level

Then, from all the blood sampling tubes, one-half of the plasma wastaken (initial baseline samples) and stored frozen at −20° C. in a 5 mlpoly(ethylene terephthalate) tube. The remaining one-half of plasma wasstored as it was at 4° C. for 20 hours and, then, stored frozen in a 5ml poly(ethylene terephthalate) tube (20-hour storage samples) in thesame manner.

Forty-eight (48) hours after blood sampling, β-TG was assayed inExamples 56 to 63 and Comparative Examples 18 to 21 and LDH and K wereassayed in Examples 64 to 72 and Comparative Examples 22 to 25. Theresults are shown in Tables 19 and 20. While β-TG is a substancecontained at a high level in the platelets and LDH and K+ are substancesoccurring at high concentrations in both the platelets and erythrocytes,these substances are released when the blood cells are stimulated ordestroyed. In the Examples wherein a sufficient amount of thecomposition was retained on the inside wall of the tube, there was nodeposition of blood cells on the inside wall so that the measured valuesafter 20 hours of storage were not much different from the initialbaseline values. In the Comparative Examples, in which small amounts ofthe clot were adhering to the inside wall, β-TG showed higher values,both initially and after 20 hours of storage, than in the Examples andLDH and K+ showed upward changes in the course of 20 hours' storage.

TABLE 20 β -TG (ng/ml) Initial Sample after 20 hr sample of standingExample 57 25 27 Example 58 25 26 Example 59 23 26 Example 60 27 25Example 61 26 23 Example 62 23 25 Example 63 23 25 Example 64 25 27Comparative Example 18 33 49 Comparative Example 19 34 51 ComparativeExample 20 32 53 Comparative Example 21 27 39

TABLE 21 LDH (IU/l) K (mEq/l) Sample after Sample after Initial 20 hrInitial 20 hr sample of standing sample of standing Example 65 130 1353.8 3.9 Example 66 125 130 3.8 3.8 Example 67 127 130 3.8 3.8 Example 68132 128 3.7 3.9 Example 69 122 126 3.8 3.8 Example 70 136 133 3.8 3.7Example 71 131 135 3.8 3.8 Example 72 135 132 3.7 3.9 ComparativeExample 22 129 156 3.8 4.8 Comparative Example 23 135 164 3.7 5.3Comparative Example 24 131 158 3.7 5.1 Comparative Example 25 133 1673.8 5.1

INDUSTRIAL APPLICABILITY

With the blood test ware of the present invention, blood coagulationfactors are rapidly activated to shorten the clotting time in asignificant degree and, at the same time, the deposition of theresulting clot on the inside wall of the ware is precluded, with theresult that separation of the serum from the clot is facilitated andcontamination of the serum with components of the clot is prevented. Theserum yield is also remarkably increased.

What is claimed is:
 1. A composition for preventing deposition of bloodcomponents to the inner surface of a blood test ware or to the surfaceof a matrix, wherein said composition comprises a blood anticoagulantand a random copolymer comprising 10 to 70 mol % of vinylprrolidone andcorrespondingly 90 to 30 mol % of vinyl acetate, said matrix comprisinga support and said composition being caused to be present on the surfaceof the inner wall of the blood test ware or of the support to preventdeposition of blood components on the surface thereof, and wherein saidinner wall or support of said matrix comprises a material selected fromthe group consisting of thermoplastic resin, thermosetting resin,modified natural resin and glass.
 2. The composition according to claim1, wherein said blood anticoagulant is at least one member selected fromthe group consisting of a salt of ethylenediaminetetraacetic acid, asalt of citric acid, a heparin salt and a salt of oxalic acid.
 3. Thecomposition according to claim 1, which further comprises an adsorbentinorganic material.
 4. The composition according to claim 3, whereinsaid adsorbent inorganic material is at least one member selected fromthe group consisting of glass, kaoline, bentonite, silica and cerite. 5.The composition according to claim 3, wherein said adsorbent inorganicmaterial has a linseed oil absorption value of 20 to 40 ml/100 g and aBET specific surface area value of 5,000 to 30,000 cm²/g.
 6. Thecomposition according to claim 3, wherein said adsorbent inorganicmaterial has a resistivity value of not larger than 1×10¹⁰ Ω.cm.
 7. Thecomposition according to claim 1, which further comprises an organicblood-coagulation-accelerating substance.
 8. The composition accordingto claim 7, wherein said organic blood-coagulation-acceleratingsubstance is ellagic acid.
 9. The composition according to claim 1,which farther comprises an antiglycolytic agent.
 10. The compositionaccording to claim 9, wherein said antiglycolytic agent is a fluoride ormannose or both.
 11. The composition according to claim 1, wherein saidblood test ware or matrix are intended for hematological, serumbiochemical or immunoserological examinations of a blood sample.
 12. Ablood test ware comprising a blood test vessel having an inner surfaceand a composition disposed on the inner surface, wherein saidcomposition comprises a blood anticoagulant and a random copolymercomprising 10 to 70 mol % of vinylprrolidone and correspondingly 90 to30 mol % of vinyl acetate.
 13. The blood test ware according to claim12, wherein said blood anticoagulant is at least one member selectedfrom the group consisting of a salt of ethylenediaminetetraacetic acid,a salt of citric acid, a heparin salt and a salt of oxalic acid.
 14. Theblood test ware according to claim 12, wherein the amount of thecomposition present on the inner surface is 1×10⁻¹⁰ to 1×⁻² g/cm². 15.The blood test ware according to claim 12, wherein the vessel comprisesa material selected from the group consisting of thermoplastic resin,thermosetting resin, modified natural resin and glass.
 16. The bloodtest ware according to claim 12, which is further comprises aserum/plasma separator.
 17. The blood test ware according to claim 12,which is intended for hematological, serum biochemical orimmunoserological examinations of a blood sample.
 18. A blood testvessel comprising a blood component deposition-preventing matrixtherein, wherein the matrix comprises a support containing a compositiondisposed on a surface thereof, and said composition comprises bloodanticoagulant and random copolymer comprising 10 to 70 mol % ofvinylpyrrolidone and correspondingly 90 to 30 mol % of vinyl acetate.19. The blood test vessel according to claim 18, wherein said bloodanticoagulant is at least one member selected from the group consistingof a salt of ethylenediaminetetraacetic acid, a salt of citric acid, aheparin salt and a salt of oxalic acid.
 20. T he blood test vesselaccording to claim 18, wherein the amount of the composition present onthe surface of the support is 1×10⁻¹⁰ to 1×10⁻² g/cm².
 21. The bloodtest according to claim 18, wherein said support comprises thermoplasticresin, thermosetting resin and modified natural resin.
 22. A method forpreventing deposition of blood components to the inner surface of ablood test ware or to the surface of a matrix, said matrix comprising asupport, which comprises causing a composition to be present on thesurface of the inner wall of the blood test ware or of the support,wherein said composition comprises a blood anticoagulant and a randomcopolymer comprising 10 to 70 mol % of vinylprrolidone andcorrespondingly 90 to 30 mol % of vinyl acetate, and said inner wall orsupport of said matrix comprises a material selected from the groupconsisting of thermoplastic resin, thermosetting resin, modified naturalresin and glass.
 23. The method according to claim 22, wherein saidblood anticoagulant is at least one member selected from the groupconsisting of a salt of ethylenediaminetetraacetic acid, a salt ofcitric acid, a heparin salt and a salt of oxalic acid.
 24. The methodaccording to claim 22, which comprises kneading said random copolymerinto a plastic batch and molding the kneaded mixture to prepare saidblood test ware or said support, to cause said random copolymer to bepresent on the surface of the inner wall of the blood test ware or ofthe support, and applying said blood anticoagulant to the inner wall ofsaid blood test ware or to the surface of said support by spray coatingor dip coating, in order to cause said composition to be present on thesurface of the inner wall of the blood test ware or of the support. 25.The method according to claim 22, which comprises, dissolving saidrandom copolymer in pure water or alcohol to obtain a solution, applyingthe solution to the inner wall of said blood test ware or to the surfaceof said support by spray coating or dip coating, and drying the coat, tocause said composition to be present on the surface of the inner wall ofthe blood test ware or of the support.
 26. The method according to claim22, which comprises dissolving or suspending said composition in amedium to obtain a solution or a suspension, applying the solution orthe suspension to the inner wall of said blood test were or to thesurface of said support by spray coating or dip coating, and drying thecoat, in order to cause said composition to be present on the surface ofthe inner wall of the blood test ware or of the support.